Antiviral compounds

ABSTRACT

The disclosure is related to anti-viral compounds, compositions containing such compounds, and therapeutic methods that include the administration of such compounds, as well as to processes and intermediates useful for preparing such compounds.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.14/541,057, filed Nov. 13, 2014, which is a continuation of U.S.application Ser. No. 14/049,133, filed Oct. 8, 2013, now U.S. Pat. No.8,921,341, which is a continuation of U.S. application Ser. No.13/783,056, filed Mar. 1, 2013, now U.S. Pat. No. 8,575,135, which is acontinuation of U.S. application Ser. No. 13/679,874, filed Nov. 16,2012, now U.S. Pat. No. 8,940,718, which application claims the benefitunder 35 U.S.C. § 119(e) of U.S. Provisional Application No. 61/560,654,filed Nov. 16, 2011, which applications are hereby incorporated byreference in their entirety.

BACKGROUND

Hepatitis C is recognized as a chronic viral disease of the liver whichis characterized by liver disease. Although drugs targeting the liverare in wide use and have shown effectiveness, toxicity and other sideeffects have limited their usefulness. Inhibitors of hepatitis C virus(HCV) are useful to limit the establishment and progression of infectionby HCV as well as in diagnostic assays for HCV.

There is a need for new HCV therapeutic agents. In particular, there isa need for HCV therapeutic agents that have broad activity against HCVgenotypes (e.g. genotypes 1a, 1b, 2a, 3a, 4a). There is also aparticular need for agents that are less susceptible to viralresistance. Resistance mutations to inhibitors have been described forHCV NS5A for genotypes 1a and 1b in Antimicrobial Agents andChemotherapy, September 2010, Volume 54, p. 3641-3650.

SUMMARY

In one embodiment the disclosure provides a compound of the disclosurewhich is a compound of formula (I):

E^(1a)-V^(1a)—C_((═O)—P) ^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

and W^(1a) is optionally substituted with one or more groupsindependently selected from halo, alkyl, haloalkyl, or cyano;

Y⁵ is —O—CH₂—, or —CH₂—O—; X⁵ is —CH₂—CH₂— or —CH═CH—;

E^(1a) is —N(H)(alkoxycarbonyl), —N(H)(cycloalkylcarbonyl) or—N(H)(cycloalkyloxycarbonyl); or E^(1a)-V^(1a) taken together areR^(9a);

E^(1b) is —N(H)(alkoxycarbonyl), —N(H)(cycloalkylcarbonyl) or—N(H)(cycloalkyloxycarbonyl); or E^(1b)-V^(1b) taken together areR^(9b);

V^(1a) and V^(1b) are each independently selected from:

P^(1a) is selected from:

P^(1b) is selected from:

and

R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.

The disclosure also provides isotopically enriched compounds that arecompounds of the disclosure that comprise an enriched isotope at one ormore positions in the compound.

The present disclosure also provides a pharmaceutical compositioncomprising a compound of the disclosure or a pharmaceutically acceptablesalt or prodrug thereof and at least one pharmaceutically acceptablecarrier.

The present disclosure also provides a pharmaceutical composition foruse in treating hepatitis C (HCV). In one embodiment the compositioncomprises at least one additional therapeutic agent for treating HCV. Inone embodiment, the therapeutic agent is selected from ribavirin, an NS3protease inhibitor, a nucleoside or nucleotide inhibitor of HCV NS5Bpolymerase, an alpha-glucosidase 1 inhibitor, a hepatoprotectant, anon-nucleoside inhibitor of HCV polymerase, or combinations thereof. Inone embodiment, composition further comprises a nucleoside or nucleotideinhibitor of HCV NSSB polymerase. In one embodiment, the nucleoside ornucleotide inhibitor of HCV NSSB polymerase is selected from ribavirin,viramidine, levovirin, a L-nucleoside, or isatoribine.

In one embodiment is provided a pharmaceutical composition comprising acompound as described herein and at least one nucleoside or nucleotideinhibitor of HCV NSSB polymerase, and at least one pharmaceuticallyacceptable carrier. In one embodiment, the composition further comprisesan interferon, a pegylated interferon, ribavirin or combinationsthereof. In one embodiment, the compound is the compound exemplified inExample PY. In one embodiment the nucleoside or nucleotide inhibitor ofHCV NS5B polymerase is sofosbuvir.

The present disclosure also provides a pharmaceutical compositionfurther comprising an interferon or pegylated interferon.

The present disclosure also provides a pharmaceutical compositionfurther comprising a nucleoside analog.

The present disclosure also provides for a pharmaceutical compositionwherein said nucleoside analogue is selected from ribavirin, viramidine,levovirin, an L-nucleoside, and isatoribine and said interferon isα-interferon or pegylated α-interferon.

The present disclosure also provides for a method of treating hepatitisC, said method comprising administering to a human patient apharmaceutical composition which comprises a therapeutically effectiveamount of a compound of the disclosure.

The present disclosure also provides a method of inhibiting HCV,comprising administering to a mammal afflicted with a conditionassociated with HCV activity, an amount of a compound of the disclosure,effective to inhibit HCV.

The present disclosure also provides a compound of the disclosure foruse in medical therapy (e.g. for use in inhibiting HCV activity ortreating a condition associated with HCV activity), as well as the useof a compound of the disclosure for the manufacture of a medicamentuseful for inhibiting HCV or the treatment of a condition associatedwith HCV activity in a mammal.

The present disclosure also provides synthetic processes and novelintermediates disclosed herein which are useful for preparing compoundsof the disclosure. Some of the compounds of the disclosure are useful toprepare other compounds of the disclosure.

In another aspect the disclosure provides a compound of the disclosure,or a pharmaceutically acceptable salt or prodrug thereof, for use in theprophylactic or therapeutic treatment of hepatitis C or a hepatitis Cassociated disorder.

In another aspect the disclosure provides a method of inhibiting HCVactivity in a sample comprising treating the sample with a compound ofthe disclosure.

Compounds of formula (I) have been found to possess useful activityagainst HCV genotypes. Additionally certain compounds of formula (I)have significant potency against resistant variants in GT1.

Accordingly, certain compounds of formula (I) possess beneficialpharmacological properties that make them well suited to fulfill thecurrent need for HCV agents with such beneficial properties.

In one embodiment the disclosure provides a compound having improvedinhibitory or pharmacokinetic properties, including enhanced activityagainst development of viral resistance, improved oral bioavailability,greater potency (for example, in inhibiting HCV activity) or extendedeffective half-life in vivo. Certain compounds of the disclosure mayhave fewer side effects, less complicated dosing schedules, or be orallyactive.

DETAILED DESCRIPTION

Reference will now be made in detail to certain embodiments of thedisclosure, examples of which are illustrated in the accompanyingstructures and formulas. While the disclosure will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the disclosure to those embodiments. Onthe contrary, the disclosure is intended to cover all alternatives,modifications, and equivalents, which may be included within the scopeof the present disclosure as defined by the embodiments.

Compounds

The compounds of the disclosure exclude compounds heretofore known.However, it is within the disclosure to use compounds that previouslywere not known to have antiviral properties for antiviral purposes (e.g.to produce an anti-viral effect in an animal). With respect to theUnited States, the compounds or compositions herein exclude compoundsthat are anticipated under 35 USC § 102 or that are obvious under 35 USC§ 103.

Whenever a compound described herein is substituted with more than oneof the same designated group, e.g., “R₁” or “A³”, then it will beunderstood that the groups may be the same or different, i.e., eachgroup is independently selected.

“Absent” Some groups are defined such that they can be absent. When agroup is absent it becomes a bond connector. The two groups that wouldotherwise be connected to that absent group are connected to each otherthrough a bond.

The “P” groups (e.g. P^(1a) and P^(1b)) defined for formula (I) hereinhave one bond to a —C(═O)— of formula (I) and one bond to a W^(1a)group. It is to be understood that a nitrogen of the P group isconnected to the —C(═O)— group of formula (I) and that a carbon of the Pgroup is connected to the W^(1a) group.

In the W^(1a) group a Y⁵ group is present. When that Y⁵ group is definedas —O—CH₂—, or —CH₂—O— group, those Y⁵ groups have a directionality. TheY⁵ group is connected to the W^(1a) group in the same left to rightdirectionality that each is drawn. So for example, when Y⁵ is —O—CH₂—,the directly following structure is intended:

For example, when Y⁵ is —CH₂—O—, the directly following structure isintended:

In the structure I, the W^(1a) group has a left-to-right directionalityas depicted in I and W^(1a) as drawn.

E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

For example, the P^(1a) group is connected to the imidazole group ofW^(1a), and the P^(1b) group is connected to the pentacyclic ring systemof W^(1a).

“Alkyl” is C₁-C₁₈ hydrocarbon containing normal, secondary, tertiary orcyclic carbon atoms. Examples are methyl (Me, —CH₃), ethyl (Et,—CH₂CH₃), 1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr,i-propyl, —CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃),2-methyl-1-propyl (i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl,—CH(CH₃)CH₂CH₃), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl(n-pentyl, —CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, and cyclopropylmethyl

“Alkenyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp² double bond. Examples include, but are not limitedto, ethylene or vinyl (—CH═CH₂), allyl (—CH₂CH═CH₂), cyclopentenyl(—C₅H₇), and 5-hexenyl (—CH₂ CH₂CH₂CH₂CH═CH₂).

“Alkynyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp triple bond. Examples include, but are not limited to,acetylenic (—CH) and propargyl (—CH₂C≡CH).

“Alkylene” refers to a saturated, branched or straight chain or cyclichydrocarbon radical of 1-18 carbon atoms, and having two monovalentradical centers derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent alkane. Typical alkyleneradicals include, but are not limited to, methylene (—CH₂—) 1,2-ethyl(—CH₂CH₂—), 1,3-propyl (—CH₂CH₂CH₂—), 1,4-butyl (—CH₂CH₂CH₂CH₂—), andthe like.

“Alkenylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkene. Typicalalkenylene radicals include, but are not limited to, 1,2-ethylene(—CH═CH—).

“Alkynylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkyne. Typicalalkynylene radicals include, but are not limited to, acetylene (—C≡C—),propargyl (—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡CH).

The term “alkoxy” or “alkyloxy,” as used herein, refers to an alkylgroup attached to the parent molecular moiety through an oxygen atom.

The term “alkoxycarbonyl,” as used herein, refers to an alkoxy groupattached to the parent molecular moiety through a carbonyl group.

The term “cycloalkyl,” as used herein, refers to a saturated monocyclic,hydrocarbon ring system having three to seven carbon atoms and zeroheteroatoms. Representative examples of cycloalkyl groups include, butare not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. Thecycloalkyl groups of the present disclosure are optionally substitutedwith one, two, three, four, or five substituents independently selectedfrom alkoxy, alkyl, aryl, cyano, halo, haloalkoxy, haloalkyl,heterocyclyl, hydroxy, hydroxyalkyl, nitro, and —NR^(x)R^(y) wherein thearyl and the heterocyclyl are further optionally substituted with one,two, or three substituents independently selected from alkoxy, alkyl,cyano, halo, haloalkoxy, haloalkyl, hydroxy, and nitro.

The term “cycloalkylcarbonyl,” as used herein, refers to a cycloalkylgroup attached to the parent molecular moiety through a carbonyl group.

The term “cycloalkyloxy,” as used herein, refers to a cycloalkyl groupattached to the parent molecular moiety through an oxygen atom.

The term “cycloalkyloxycarbonyl,” as used herein, refers to acycloalkyloxy group attached to the parent molecular moiety through acarbonyl group.

“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbonatoms derived by the removal of one hydrogen atom from a single carbonatom of a parent aromatic ring system. Typical aryl groups include, butare not limited to, radicals derived from benzene, substituted benzene,naphthalene, anthracene, biphenyl, and the like.

“Arylalkyl” refers to an acyclic alkyl radical in which one of thehydrogen atoms bonded to a carbon atom, typically a terminal or spacarbon atom, is replaced with an aryl radical. Typical arylalkyl groupsinclude, but are not limited to, benzyl, 2-phenylethan-1-yl,naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. The arylalkyl group comprises 6to 20 carbon atoms, e.g., the alkyl moiety, including alkanyl, alkenylor alkynyl groups, of the arylalkyl group is 1 to 6 carbon atoms and thearyl moiety is 5 to 14 carbon atoms.

“Substituted alkyl”, “substituted aryl”, and “substituted arylalkyl”mean alkyl, aryl, and arylalkyl respectively, in which one or morehydrogen atoms are each independently replaced with a non-hydrogensubstituent. Typical substituents include, but are not limited to: halo(e.g. F, Cl, Br, I), —R, —OR, —SR, —NR₂, —CF₃, —CCl₃, —OCF₃, —CN, —NO₂,—N(R)C(═O)R, —C(═O)R, —OC(═O)R, —C(O)OR, —C(═O)NRR, —S(═O)R, —S(═O)₂OR,—S(═O)₂R, —OS(═O)₂OR, —S(═O)₂NRR, and each R is independently —H, alkyl,aryl, arylalkyl, or heterocycle. Alkylene, alkenylene, and alkynylenegroups may also be similarly substituted.

The term “optionally substituted” in reference to a particular moiety ofthe compound of formula I, (e.g., an optionally substituted aryl group)refers to a moiety having 0, 1, 2, or more substituents.

The symbol “

” in a ring structure means that a bond is a single or double bond. In anon-limiting example,

can be

“Haloalkyl” as used herein includes an alkyl group substituted with oneor more halogens (e.g. F, Cl, Br, or I). Representative examples ofhaloalkyl include trifluoromethyl, 2,2,2-trifluoroethyl, and2,2,2-trifluoro-1-(trifluoromethyl)ethyl.

“Heterocycle” or “heterocyclyl” as used herein includes by way ofexample and not limitation these heterocycles described in Paquette, LeoA.; Principles of Modern Heterocyclic Chemistry (W.A. Benjamin, NewYork, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; The Chemistryof Heterocyclic Compounds, A Series of Monographs” (John Wiley & Sons,New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and28; and J. Am. Chem. Soc. (1960) 82:5566. In one specific embodiment,“heterocycle” includes a “carbocycle” as defined herein, wherein one ormore (e.g. 1, 2, 3, or 4) carbon atoms have been replaced with aheteroatom (e.g. O, N, or S). The term heterocycle also includes“heteroaryl” which is a heterocycle wherein at least one heterocyclicrings is aromatic.

Examples of heterocycles include by way of example and not limitationpyridyl, dihydropyridyl, tetrahydropyridyl (piperidyl), thiazolyl,tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl, pyrimidinyl,furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl,benzofuranyl, thianaphthalenyl, indolyl, indolenyl, quinolinyl,isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl,2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl,azocinyl, triazinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl,thienyl, thianthrenyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl,phenoxathinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl,pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, 1H-indazolyl, purinyl,4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl,quinazolinyl, cinnolinyl, pteridinyl, 4H-carbazolyl, carbazolyl,β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl,chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl,oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl,isatinoyl, and bis-tetrahydrofuranyl:

By way of example and not limitation, carbon bonded heterocycles arebonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2,3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan,tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole,position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4,or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of anaziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6,7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of anisoquinoline. Still more typically, carbon bonded heterocycles include2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl,4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl,4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl,5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

By way of example and not limitation, nitrogen bonded heterocycles arebonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine,2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline,3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline,piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of aisoindole, or isoindoline, position 4 of a morpholine, and position 9 ofa carbazole, or β-carboline. Still more typically, nitrogen bondedheterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl, 1-imidazolyl,1-pyrazolyl, and 1-piperidinyl.

“Carbocycle” refers to a saturated, unsaturated or aromatic ring havingup to about 25 carbon atoms. Typically, a carbocycle has about 3 to 7carbon atoms as a monocycle, about 7 to 12 carbon atoms as a bicycle,and up to about 25 carbon atoms as a polycycle. Monocyclic carbocyclestypically have 3 to 6 ring atoms, still more typically 5 or 6 ringatoms. Bicyclic carbocycles typically have 7 to 12 ring atoms, e.g.,arranged as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10ring atoms arranged as a bicyclo [5,6] or [6,6] system. The termcarbocycle includes “cycloalkyl” which is a saturated or unsaturatedcarbocycle. Examples of monocyclic carbocycles include cyclopropyl,cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl,1-cyclohex-3-enyl, phenyl, spiryl and naphthyl.

The term “amino,” as used herein, refers to —NH₂.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g., melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

The term “treatment” or “treating,” to the extent it relates to adisease or condition includes preventing the disease or condition fromoccurring, inhibiting the disease or condition, eliminating the diseaseor condition, and/or relieving one or more symptoms of the disease orcondition.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork. Many organic compounds exist in optically active forms, i.e., theyhave the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes (D and L) or (Rand S) are used to denote the absolute configuration of the moleculeabout its chiral center(s). The prefixes d and 1 or (+) and (−) areemployed to designate the sign of rotation of plane-polarized light bythe compound, with (−) or 1 meaning that the compound is levorotatory. Acompound prefixed with (+) or d is dextrorotatory. For a given chemicalstructure, these stereoisomers are identical except that they are mirrorimages of one another. A specific stereoisomer may also be referred toas an enantiomer, and a mixture of such isomers is often called anenantiomeric mixture. A 50:50 mixture of enantiomers is referred to as aracemic mixture or a racemate, which may occur where there has been nostereoselection or stereospecificity in a chemical reaction or process.The terms “racemic mixture” and “racemate” refer to an equimolar mixtureof two enantiomeric species, devoid of optical activity. The disclosureincludes all stereoisomers of the compounds described herein.

Prodrugs

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates a compound of thedisclosure that inhibits HCV activity (“the active inhibitorycompound”). The compound may be formed from the prodrug as a result of:(i) spontaneous chemical reaction(s), (ii) enzyme catalyzed chemicalreaction(s), (iii) photolysis, and/or (iv) metabolic chemicalreaction(s).

“Prodrug moiety” refers to a labile functional group which separatesfrom the active inhibitory compound during metabolism, systemically,inside a cell, by hydrolysis, enzymatic cleavage, or by some otherprocess (Bundgaard, Hans, “Design and Application of Prodrugs” in ATextbook of Drug Design and Development (1991), P. Krogsgaard-Larsen andH. Bundgaard, Eds. Harwood Academic Publishers, pp. 113-191). Enzymeswhich are capable of an enzymatic activation mechanism with the prodrugcompounds of the disclosure include, but are not limited to, amidases,esterases, microbial enzymes, phospholipases, cholinesterases, andphosphases. Prodrug moieties can serve to enhance solubility, absorptionand lipophilicity to optimize drug delivery, bioavailability andefficacy. A prodrug moiety may include an active metabolite or drugitself.

Exemplary prodrug moieties include the hydrolytically sensitive orlabile acyloxymethyl esters CH₂OC(═O)R⁹⁹ and acyloxymethyl carbonatesCH₂OC(═O)OR⁹⁹ where R⁹⁹ is C₁-C₆ alkyl, C₁-C₆ substituted alkyl, C₆-C₂₀aryl or C₆-C₂₀ substituted aryl. The acyloxyalkyl ester was first usedas a prodrug strategy for carboxylic acids and then applied tophosphates and phosphonates by Farquhar et al. (1983) J. Pharm. Sci. 72:324; also U.S. Pat. Nos. 4,816,570, 4,968,788, 5,663,159 and 5,792,756.Subsequently, the acyloxyalkyl ester was used to deliver phosphonicacids across cell membranes and to enhance oral bioavailability. A closevariant of the acyloxyalkyl ester, the alkoxycarbonyloxyalkyl ester(carbonate), may also enhance oral bioavailability as a prodrug moietyin the compounds of the combinations of the disclosure. An exemplaryacyloxymethyl ester is pivaloyloxymethoxy, (POM) CH₂OC(═O)C(CH₃)₃. Anexemplary acyloxymethyl carbonate prodrug moiety ispivaloyloxymethylcarbonate (POC) CH₂OC(═O)OC(CH₃)₃.

Aryl esters of phosphorus groups, especially phenyl esters, are reportedto enhance oral bioavailability (De Lombaert et al. (1994) J. Med. Chem.37: 498). Phenyl esters containing a carboxylic ester ortho to aphosphate have also been described (Khamnei and Torrence, (1996) J. Med.Chem. 39:4109-4115). Benzyl esters are reported to generate parentphosphonic acids. In some cases, substituents at the ortho- orpara-position may accelerate the hydrolysis. Benzyl analogs with anacylated phenol or an alkylated phenol may generate the phenoliccompound through the action of enzymes, e.g., esterases, oxidases, etc.,which in turn undergoes cleavage at the benzylic C—O bond to generatephosphoric acid and a quinone methide intermediate. Examples of thisclass of prodrugs are described by Mitchell et al. (1992) J. Chem. Soc.Perkin Trans. II 2345; Glazier WO 91/19721. Still other benzylicprodrugs have been described containing a carboxylic ester-containinggroup attached to the benzylic methylene (Glazier WO 91/19721).Thio-containing prodrugs are reported to be useful for the intracellulardelivery of phosphonate drugs. These proesters contain an ethylthiogroup in which the thiol group is either esterified with an acyl groupor combined with another thiol group to form a disulfide.Deesterification or reduction of the disulfide generates the free thiointermediate which subsequently breaks down to the phosphoric acid andepisulfide (Puech et al. (1993) Antiviral Res., 22: 155-174; Benzaria etal. (1996) J. Med. Chem. 39: 4958).

Protecting Groups

In the context of the present disclosure, protecting groups includeprodrug moieties and chemical protecting groups.

“Protecting group” refers to a moiety of a compound that masks or altersthe properties of a functional group or the properties of the compoundas a whole. Chemical protecting groups and strategies forprotection/deprotection are well known in the art. See e.g., ProtectiveGroups in Organic Chemistry, Theodora W. Greene, John Wiley & Sons,Inc., New York, 1991. Protecting groups are often utilized to mask thereactivity of certain functional groups, to assist in the efficiency ofdesired chemical reactions, e.g., making and breaking chemical bonds inan ordered and planned fashion. Protection of functional groups of acompound alters other physical properties besides the reactivity of theprotected functional group, such as the polarity, lipophilicity(hydrophobicity), and other properties which can be measured by commonanalytical tools. Chemically protected intermediates may themselves bebiologically active or inactive.

Protected compounds may also exhibit altered, and in some cases,optimized properties in vitro and in vivo, such as passage throughcellular membranes and resistance to enzymatic degradation orsequestration. In this role, protected compounds with intendedtherapeutic effects may be referred to as prodrugs. Another function ofa protecting group is to convert the parental drug into a prodrug,whereby the parental drug is released upon conversion of the prodrug invivo. Because active prodrugs may be absorbed more effectively than theparental drug, prodrugs may possess greater potency in vivo than theparental drug. Protecting groups are removed either in vitro, in theinstance of chemical intermediates, or in vivo, in the case of prodrugs.With chemical intermediates, it is not particularly important that theresulting products after deprotection, e.g., alcohols, bephysiologically acceptable, although in general it is more desirable ifthe products are pharmacologically innocuous.

Protecting groups are available, commonly known and used, and areoptionally used to prevent side reactions with the protected groupduring synthetic procedures, i.e. routes or methods to prepare thecompounds of the disclosure. For the most part the decision as to whichgroups to protect, when to do so, and the nature of the chemicalprotecting group “PG” will be dependent upon the chemistry of thereaction to be protected against (e.g., acidic, basic, oxidative,reductive or other conditions) and the intended direction of thesynthesis.

PGs do not need to be, and generally are not, the same if the compoundis substituted with multiple PGs. In general, PG will be used to protectfunctional groups such as carboxyl, hydroxyl, thio, or amino groups andto thus prevent side reactions or to otherwise facilitate the syntheticefficiency. The order of deprotection to yield free deprotected groupsis dependent upon the intended direction of the synthesis and thereaction conditions to be encountered, and may occur in any order asdetermined by the artisan.

Various functional groups of the compounds of the disclosure may beprotected. For example, protecting groups for OH groups (whetherhydroxyl, carboxylic acid, phosphonic acid, or other functions) include“ether- or ester-forming groups”. Ether- or ester-forming groups arecapable of functioning as chemical protecting groups in the syntheticschemes set forth herein. However, some hydroxyl and thio protectinggroups are neither ether-nor ester-forming groups, as will be understoodby those skilled in the art, and are included with amides, discussedbelow.

A very large number of hydroxyl protecting groups and amide-forminggroups and corresponding chemical cleavage reactions are described inProtective Groups in Organic Synthesis, Theodora W. Greene (John Wiley &Sons, Inc., New York, 1991, ISBN 0-471-62301-6) (“Greene”). See alsoKocienski, Philip J.; Protecting Groups (Georg Thieme Verlag Stuttgart,New York, 1994), which is incorporated by reference in its entiretyherein. In particular Chapter 1, Protecting Groups: An Overview, pages1-20, Chapter 2, Hydroxyl Protecting Groups, pages 21-94, Chapter 3,Diol Protecting Groups, pages 95-117, Chapter 4, Carboxyl ProtectingGroups, pages 118-154, Chapter 5, Carbonyl Protecting Groups, pages155-184. For protecting groups for carboxylic acid, phosphonic acid,phosphonate, sulfonic acid and other protecting groups for acids seeGreene as set forth below.

By way of example and not limitation, variables described herein mayrecursive substituents in certain embodiments. Typically, each of thesemay independently occur 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9,8, 7, 6, 5, 4, 3, 2, 1, or 0, times in a given embodiment. Moretypically, each of these may independently occur 12 or fewer times in agiven embodiment. Whenever a compound described herein is substitutedwith more than one of the same designated group, e.g., “R¹” or “R³”,then it will be understood that the groups may be the same or different,i.e., each group is independently selected. Wavy lines indicate the siteof covalent bond attachments to the adjoining groups, moieties, oratoms.

In one embodiment of the disclosure, the compound is in an isolated andpurified form. Generally, the term “isolated and purified” means thatthe compound is substantially free from biological materials (e.g.blood, tissue, cells, etc.). In one specific embodiment of thedisclosure, the term means that the compound or conjugate of thedisclosure is at least about 50 wt. % free from biological materials; inanother specific embodiment, the term means that the compound orconjugate of the disclosure is at least about 75 wt. % free frombiological materials; in another specific embodiment, the term meansthat the compound or conjugate of the disclosure is at least about 90wt. % free from biological materials; in another specific embodiment,the term means that the compound or conjugate of the disclosure is atleast about 98 wt. % free from biological materials; and in anotherembodiment, the term means that the compound or conjugate of thedisclosure is at least about 99 wt. % free from biological materials. Inanother specific embodiment, the disclosure provides a compound orconjugate of the disclosure that has been synthetically prepared (e.g.,ex vivo).

Stereoisomers

The compounds of the disclosure may have chiral centers, e.g., chiralcarbon or phosphorus atoms. The compounds of the disclosure thus includeracemic mixtures of all stereoisomers, including enantiomers,diastereomers, and atropisomers. In addition, the compounds of thedisclosure include enriched or resolved optical isomers at any or allasymmetric, chiral atoms. In other words, the chiral centers apparentfrom the depictions are provided as the chiral isomers or racemicmixtures. Both racemic and diastereomeric mixtures, as well as theindividual optical isomers isolated or synthesized, substantially freeof their enantiomeric or diastereomeric partners, are all within thescope of the disclosure. The racemic mixtures are separated into theirindividual, substantially optically pure isomers through well-knowntechniques such as, for example, the separation of diastereomeric saltsformed with optically active adjuncts, e.g., acids or bases followed byconversion back to the optically active substances. In most instances,the desired optical isomer is synthesized by means of stereospecificreactions, beginning with the appropriate stereoisomer of the desiredstarting material.

The compounds of the disclosure can also exist as tautomeric isomers incertain cases. Although only one tautomer may be depicted, all suchforms are contemplated within the scope of the disclosure. For example,ene-amine tautomers can exist for purine, pyrimidine, imidazole,guanidine, amidine, and tetrazole systems and all their possibletautomeric forms are within the scope of the disclosure.

Salts and Hydrates

Examples of physiologically or pharmaceutically acceptable salts of thecompounds of the disclosure include salts derived from an appropriatebase, such as an alkali metal (for example, sodium), an alkaline earthmetal (for example, magnesium), ammonium and NX₄ ⁺ (wherein X is C₁-C₄alkyl). Physiologically acceptable salts of a hydrogen atom or an aminogroup include salts of organic carboxylic acids such as acetic, benzoic,lactic, fumaric, tartaric, maleic, malonic, malic, isethionic,lactobionic and succinic acids; organic sulfonic acids, such asmethanesulfonic, ethanesulfonic, benzenesulfonic and p-toluenesulfonicacids; and inorganic acids, such as hydrochloric, sulfuric, phosphoricand sulfamic acids. Physiologically acceptable salts of a compound of ahydroxy group include the anion of said compound in combination with asuitable cation such as Na⁺ and NX₄ ⁺ (wherein X is independentlyselected from H or a C₁-C₄ alkyl group).

For therapeutic use, salts of active ingredients of the compounds of thedisclosure will typically be physiologically acceptable, i.e. they willbe salts derived from a physiologically acceptable acid or base.However, salts of acids or bases which are not physiologicallyacceptable may also find use, for example, in the preparation orpurification of a physiologically acceptable compound. All salts,whether or not derived from a physiologically acceptable acid or base,are within the scope of the present disclosure.

Metal salts typically are prepared by reacting the metal hydroxide witha compound of this disclosure. Examples of metal salts which areprepared in this way are salts containing Li⁺, Na⁺, and K⁺. A lesssoluble metal salt can be precipitated from the solution of a moresoluble salt by addition of the suitable metal compound.

In addition, salts may be formed from acid addition of certain organicand inorganic acids, e.g., HCl, HBr, H₂SO₄, H₃PO₄ or organic sulfonicacids, to basic centers, typically amines, or to acidic groups. Finally,it is to be understood that the compositions herein comprise compoundsof the disclosure in their un-ionized, as well as zwitterionic form, andcombinations with stoichiometric amounts of water as in hydrates.

Also included within the scope of this disclosure are the salts of theparental compounds with one or more amino acids. Any of the natural orunnatural amino acids are suitable, especially the naturally-occurringamino acids found as protein components, although the amino acidtypically is one bearing a side chain with a basic or acidic group,e.g., lysine, arginine or glutamic acid, or a neutral group such asglycine, serine, threonine, alanine, isoleucine, or leucine.

Methods of Inhibition of HCV

Another aspect of the disclosure relates to methods of inhibiting theactivity of HCV comprising the step of treating a sample suspected ofcontaining HCV with a compound or composition of the disclosure.

Compounds of the disclosure may act as inhibitors of HCV, asintermediates for such inhibitors or have other utilities as describedbelow. The inhibitors will generally bind to locations on the surface orin a cavity of the liver. Compounds binding in the liver may bind withvarying degrees of reversibility. Those compounds binding substantiallyirreversibly are ideal candidates for use in this method of thedisclosure. Once labeled, the substantially irreversibly bindingcompounds are useful as probes for the detection of HCV. Accordingly,the disclosure relates to methods of detecting NS3 in a sample suspectedof containing HCV comprising the steps of: treating a sample suspectedof containing HCV with a composition comprising a compound of thedisclosure bound to a label; and observing the effect of the sample onthe activity of the label. Suitable labels are well known in thediagnostics field and include stable free radicals, fluorophores,radioisotopes, enzymes, chemiluminescent groups and chromogens. Thecompounds herein are labeled in conventional fashion using functionalgroups such as hydroxyl or amino. In one embodiment the disclosureprovides a compound of any one of formulae (I) and (A1)-(A4) thatcomprises or that is bound or linked to one or more detectable labels.Within the context of the disclosure samples suspected of containing HCVinclude natural or man-made materials such as living organisms; tissueor cell cultures; biological samples such as biological material samples(blood, serum, urine, cerebrospinal fluid, tears, sputum, saliva, tissuesamples, and the like); laboratory samples; food, water, or air samples;bioproduct samples such as extracts of cells, particularly recombinantcells synthesizing a desired glycoprotein; and the like. Typically thesample will be suspected of containing HCV. Samples can be contained inany medium including water and organic solvent/water mixtures. Samplesinclude living organisms such as humans, and man made materials such ascell cultures.

The treating step of the disclosure comprises adding the compound of thedisclosure to the sample or it comprises adding a precursor of thecomposition to the sample. The addition step comprises any method ofadministration as described above.

If desired, the activity of HCV after application of the compound can beobserved by any method including direct and indirect methods ofdetecting HCV activity. Quantitative, qualitative, and semiquantitativemethods of determining HCV activity are all contemplated. Typically oneof the screening methods described above are applied, however, any othermethod such as observation of the physiological properties of a livingorganism are also applicable.

Many organisms contain HCV. The compounds of this disclosure are usefulin the treatment or prophylaxis of conditions associated with HCVactivation in animals or in man.

However, in screening compounds capable of inhibiting HCV activity itshould be kept in mind that the results of enzyme assays may not alwayscorrelate with cell culture assays. Thus, a cell based assay shouldtypically be the primary screening tool.

Pharmaceutical Formulations

The compounds of this disclosure are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic. All formulations will optionally contain excipientssuch as those set forth in the Handbook of Pharmaceutical Excipients(1986). Excipients include ascorbic acid and other antioxidants,chelating agents such as EDTA, carbohydrates such as dextrin,hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and thelike. The pH of the formulations ranges from about 3 to about 11, but isordinarily about 7 to 10. Typically, the compound will be administeredin a dose from 0.01 milligrams to 2 grams. In one embodiment, the dosewill be from about 10 milligrams to 450 milligrams. In anotherembodiment, the dosage will be from about 25 to about 250 milligrams. Inanother embodiment, the dosage will be about 50 or 100 milligrams. Inone embodiment, the dosage will be about 100 milligrams. It iscontemplated that the compound may be administered once, twice or threetimes a day.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations, both for veterinary and for human use, of the disclosurecomprise at least one active ingredient, as above defined, together withone or more acceptable carriers therefor and optionally othertherapeutic ingredients. The carrier(s) must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand physiologically innocuous to the recipient thereof.

The formulations include those suitable for the foregoing administrationroutes. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Techniques and formulations generally are found in Remington'sPharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methodsinclude the step of bringing into association the active ingredient withthe carrier which constitutes one or more accessory ingredients. Ingeneral the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product.

Formulations of the present disclosure suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or a suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas a powder or granules, optionally mixed with a binder, lubricant,inert diluent, preservative, surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered active ingredient moistened with an inert liquid diluent. Thetablets may optionally be coated or scored and optionally are formulatedso as to provide slow or controlled release of the active ingredienttherefrom.

For administration to the eye or other external tissues e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w (including active ingredient(s) in a range between 0.1%and 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.),preferably 0.2 to 15% w/w and most preferably 0.5 to 10% w/w. Whenformulated in an ointment, the active ingredients may be employed witheither a paraffinic or a water-miscible ointment base. Alternatively,the active ingredients may be formulated in a cream with an oil-in-watercream base.

If desired, the aqueous phase of the cream base may include, forexample, at least 30% w/w of a polyhydric alcohol, i.e. an alcoholhaving two or more hydroxyl groups such as propylene glycol, butane1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol(including PEG 400) and mixtures thereof. The topical formulations maydesirably include a compound which enhances absorption or penetration ofthe active ingredient through the skin or other affected areas. Examplesof such dermal penetration enhancers include dimethyl sulphoxide andrelated analogs.

The oily phase of the emulsions of this disclosure may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the disclosure include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties. The cream should preferablybe a non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters known asCrodamol CAP may be used, the last three being preferred esters. Thesemay be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical formulations according to the present disclosure compriseone or more compounds of the disclosure together with one or morepharmaceutically acceptable carriers or excipients and optionally othertherapeutic agents. Pharmaceutical formulations containing the activeingredient may be in any form suitable for the intended method ofadministration. When used for oral use for example, tablets, troches,lozenges, aqueous or oil suspensions, dispersible powders or granules,emulsions, hard or soft capsules, syrups or elixirs may be prepared.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsincluding sweetening agents, flavoring agents, coloring agents andpreserving agents, in order to provide a palatable preparation. Tabletscontaining the active ingredient in admixture with non-toxicpharmaceutically acceptable excipient which are suitable for manufactureof tablets are acceptable. These excipients may be, for example, inertdiluents, such as calcium or sodium carbonate, lactose, lactosemonohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the disclosure contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcelluose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia,and dispersing or wetting agents such as a naturally occurringphosphatide (e.g., lecithin), a condensation product of an alkyleneoxide with a fatty acid (e.g., polyoxyethylene stearate), a condensationproduct of ethylene oxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by the addition of an antioxidant such asascorbic acid.

Dispersible powders and granules of the disclosure suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the disclosure may also be in theform of oil-in-water emulsions. The oily phase may be a vegetable oil,such as olive oil or arachis oil, a mineral oil, such as liquidparaffin, or a mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion may also contain sweetening and flavoring agents. Syrups andelixirs may be formulated with sweetening agents, such as glycerol,sorbitol or sucrose. Such formulations may also contain a demulcent, apreservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the disclosure may be in the form ofa sterile injectable preparation, such as a sterile injectable aqueousor oleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye dropswherein the active ingredient is dissolved or suspended in a suitablecarrier, especially an aqueous solvent for the active ingredient. Theactive ingredient is preferably present in such formulations in aconcentration of 0.5 to 20%, advantageously 0.5 to 10% particularlyabout 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in increments suchas 0.5 microns, 1 micron, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration may beprepared according to conventional methods and may be delivered withother therapeutic agents such as compounds heretofore used in thetreatment or prophylaxis of conditions associated with HCV activity.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

The formulations are presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this disclosure may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

The disclosure further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefor.

Veterinary carriers are materials useful for the purpose ofadministering the composition and may be solid, liquid or gaseousmaterials which are otherwise inert or acceptable in the veterinary artand are compatible with the active ingredient. These veterinarycompositions may be administered orally, parenterally or by any otherdesired route.

Compounds of the disclosure can also be formulated to provide controlledrelease of the active ingredient to allow less frequent dosing or toimprove the pharmacokinetic or toxicity profile of the activeingredient. Accordingly, the disclosure also provides compositionscomprising one or more compounds of the disclosure formulated forsustained or controlled release.

Effective dose of active ingredient depends at least on the nature ofthe condition being treated, toxicity, whether the compound is beingused prophylactically (lower doses), the method of delivery, and thepharmaceutical formulation, and will be determined by the clinicianusing conventional dose escalation studies.

Routes of Administration

One or more compounds of the disclosure (herein referred to as theactive ingredients) are administered by any route appropriate to thecondition to be treated. Suitable routes include oral, rectal, nasal,topical (including buccal and sublingual), vaginal and parenteral(including subcutaneous, intramuscular, intravenous, intradermal,intrathecal and epidural), and the like. It will be appreciated that thepreferred route may vary with for example the condition of therecipient. An advantage of the compounds of this disclosure is that theyare orally bioavailable and can be dosed orally.

HCV Combination Therapy

In another embodiment, non-limiting examples of suitable combinationsinclude combinations of one or more compounds of formula (I) and (A1-A4)with one or more interferons, ribavirin or its analogs, HCV NS3 proteaseinhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants,nucleoside or nucleotide inhibitors of HCV NS5B polymerase,non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors,TLR-7 agonists, cyclophillin inhibitors, HCV IRES inhibitors,pharmacokinetic enhancers, and other drugs or therapeutic agents fortreating HCV.

More specifically, one or more compounds as described herein may becombined with one or more compounds selected from the group consistingof:

1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron), pegylatedrIFN-alpha 2a (Pegasys), rIFN-alpha 2b (Intron A), rIFN-alpha 2a(Roferon-A), interferon alpha (MOR-22, OPC-18, Alfaferone, Alfanative,Multiferon, subalin), interferon alfacon-1 (Infergen), interferonalpha-n1 (Wellferon), interferon alpha-n3 (Alferon), interferon-beta(Avonex, DL-8234), interferon-omega (omega DUROS, Biomed 510),albinterferon alpha-2b (Albuferon), IFN alpha-2b XL, BLX-883 (Locteron),DA-3021, glycosylated interferon alpha-2b (AVI-005), PEG-Infergen,PEGylated interferon lambda-1 (PEGylated IL-29), and belerofon;

2) ribavirin and its analogs, e.g., ribavirin (Rebetol, Copegus), andtaribavirin (Viramidine);

3) HCV NS3 protease inhibitors, e.g., boceprevir (SCH-503034, SCH-7),telaprevir (VX-950), TMC435350, BI-1335, BI-1230, MK-7009, VBY-376,VX-500, GS-9256, GS-9451, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530,YH5531, ABT-450, ACH-1625, ITMN-191, MK5172, MK6325, and MK2748;

4) alpha-glucosidase 1 inhibitors, e.g., celgosivir (MX-3253), Miglitol,and UT-231B;

5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738, GS-9450(LB-84451), silibilin, and MitoQ;

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, e.g.,R1626, R7128 (R4048), IDX184, IDX-102, BCX-4678, valopicitabine(NM-283), MK-0608, sofosbuvir (GS-7977 (formerly PSI-7977)), and INX-189(now BMS986094);

7) non-nucleoside inhibitors of HCV NSSB polymerase, e.g., PF-868554,VCH-759, VCH-916, JTK-652, MK-3281, GS-9190, VBY-708, VCH-222, A848837,ANA-598, GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796(nesbuvir), GSK625433, BILN-1941, XTL-2125, ABT-072, ABT-333, GS-9669,PSI-7792, and GS-9190;

8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), BMS-790052, ACH-3102,ACH-2928, MK8325, MK4882, MK8742, PSI-461, IDX719, and A-689;

9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773, ANA-975,AZD-8848 (DSP-3025), and SM-360320;

10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and NIM811; 11)HCV IRES inhibitors, e.g., MCI-067;

12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452, PF-4194477,TMC-41629, GS-9350, GS-9585, and roxythromycin; and

13) other drugs for treating HCV, e.g., thymosin alpha 1 (Zadaxin),nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir, GI-5005,XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOV-205, tarvacin,EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032, BMS-791325,Bavituximab, MDX-1106 (ONO-4538), Oglufanide, and VX-497 (merimepodib).

More specifically, one or more compounds as described herein may becombined with one or more compounds selected from the group consistingof non-nucleoside inhibitors of HCV NSSB polymerase (ABT-072 andABT-333), HCV NS5A inhibitors (ACH-3102 and ACH-2928) and HCV NS3protease inhibitors (ABT-450 and ACH-1625).

In yet another embodiment, the present application disclosespharmaceutical compositions comprising a compound as described herein,or a pharmaceutically acceptable salt, solvate, and/or ester thereof, incombination with at least one additional therapeutic agent, and apharmaceutically acceptable carrier or excipient.

According to one embodiment, the therapeutic agent used in combinationwith the compound as described herein can be any agent having atherapeutic effect when used in combination with the compound asdescribed herein. For example, the therapeutic agent used in combinationwith the compound as described herein can be interferons, ribavirinanalogs, NS3 protease inhibitors, NS5b polymerase inhibitors,alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleosideinhibitors of HCV, and other drugs for treating HCV.

In another embodiment, the present application provides pharmaceuticalcompositions comprising a compound of formula (I), or a pharmaceuticallyacceptable salt, solvate, and/or ester thereof, in combination with atleast one additional therapeutic agent selected from the groupconsisting of pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a,rIFN-alpha 2b, IFN alpha-2b XL, rIFN-alpha 2a, consensus IFN alpha,infergen, rebif, locteron, AVI-005, PEG-infergen, pegylated IFN-beta,oral interferon alpha, feron, reaferon, intermax alpha, r-IFN-beta,infergen+actimmune, IFN-omega with DUROS, albuferon, rebetol, copegus,levovirin, VX-497, viramidine (taribavirin), A-831, A-689, NM-283,valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB 1941, MK-0608,NM-107, R7128, VCH-759, PF-868554, GSK625433, XTL-2125, SCH-503034(SCH-7), VX-950 (Telaprevir), ITMN-191, and BILN-2065, MX-3253(celgosivir), UT-231B, IDN-6556, ME 3738, MitoQ, and LB-84451,benzimidazole derivatives, benzo-1,2,4-thiadiazine derivatives, andphenylalanine derivatives, zadaxin, nitazoxanide (alinea), BIVN-401(virostat), DEBIO-025, VGX-410C, EMZ-702, AVI 4065, bavituximab,oglufanide, PYN-17, KPE02003002, actilon (CPG-10101), KRN-7000, civacir,GI-5005, ANA-975 (isatoribine), XTL-6865, ANA 971, NOV-205, tarvacin,EHC-18, and NIM811 and a pharmaceutically acceptable carrier orexcipient.

In yet another embodiment, the present application provides acombination pharmaceutical agent comprising:

a) a first pharmaceutical composition comprising a compound of formula(I), or a pharmaceutically acceptable salt, solvate, or ester thereof;and

b) a second pharmaceutical composition comprising at least oneadditional therapeutic agent selected from the group consisting of HIVprotease inhibiting compounds, HIV non-nucleoside inhibitors of reversetranscriptase, HIV nucleoside inhibitors of reverse transcriptase, HIVnucleotide inhibitors of reverse transcriptase, HIV integraseinhibitors, gp41 inhibitors, CXCR4 inhibitors, gp120 inhibitors, CCR5inhibitors, interferons, ribavirin analogs, NS3 protease inhibitors,alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleosideinhibitors of HCV, and other drugs for treating HCV, and combinationsthereof.

In another embodiment is provided a pharmaceutical compositioncomprising a compound of formula (I) as described herein and anucleoside or nucleotide inhibitors of HCV NSSB polymerase andoptionally an interferon or ribavirin. In one embodiment, the compoundis methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatehaving the formula:

and the inhibitor is sofosbuvir.

Combinations of the compounds of formula I and additional activetherapeutic agents may be selected to treat patients infected with HCVand other conditions such as HIV infections. Accordingly, the compoundsof formula I may be combined with one or more compounds useful intreating HIV, for example HIV protease inhibiting compounds,non-nucleoside inhibitors of HIV reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4inhibitors, gp120 inhibitors, CCR5 inhibitors, interferons, ribavirinanalogs, NS3 protease inhibitors, NS5b polymerase inhibitors,alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleosideinhibitors of HCV, and other drugs for treating HCV.

More specifically, one or more compounds of formula (I) or (A1)-(A4) maybe combined with one or more compounds selected from the groupconsisting of 1) HIV protease inhibitors, e.g., amprenavir, atazanavir,fosamprenavir, indinavir, lopinavir, ritonavir, lopinavir+ritonavir,nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126,TMC-114, mozenavir (DMP-450), JE-2147 (AG1776), AG1859, DG35, L-756423,R00334649, KNI-272, DPC-681, DPC-684, and GW640385X, DG17, PPL-100, 2) aHIV non-nucleoside inhibitor of reverse transcriptase, e.g.,capravirine, emivirine, delaviridine, efavirenz, nevirapine, (+)calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963, MIV-150,and TMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355 BS, VRX 840773,UK-453,061, RDEA806, 3) a HIV nucleoside inhibitor of reversetranscriptase, e.g., zidovudine, emtricitabine, didanosine, stavudine,zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine, alovudine,MIV-210, racivir (±-FTC), D-d4FC, emtricitabine, phosphazide, fozivudinetidoxil, fosalvudine tidoxil, apricitibine (AVX754), amdoxovir, KP-1461,abacavir+lamivudine, abacavir+lamivudine+zidovudine,zidovudine+lamivudine, 4) a HIV nucleotide inhibitor of reversetranscriptase, e.g., tenofovir, tenofovir disoproxilfumarate+emtricitabine, tenofovir disoproxilfumarate+emtricitabine+efavirenz, and adefovir, 5) a HIV integraseinhibitor, e.g., curcumin, derivatives of curcumin, chicoric acid,derivatives of chicoric acid, 3,5-dicaffeoylquinic acid, derivatives of3,5-dicaffeoylquinic acid, aurintricarboxylic acid, derivatives ofaurintricarboxylic acid, caffeic acid phenethyl ester, derivatives ofcaffeic acid phenethyl ester, tyrphostin, derivatives of tyrphostin,quercetin, derivatives of quercetin, S-1360, zintevir (AR-177),L-870812, and L-870810, MK-0518 (raltegravir), BMS-707035, MK-2048,BA-011, BMS-538158, GSK364735C, 6) a gp41 inhibitor, e.g., enfuvirtide,sifuvirtide, FB006M, TRI-1144, SPC3, DES6, Locus gp41, CovX, and REP 9,7) a CXCR4 inhibitor, e.g., AMD-070, 8) an entry inhibitor, e.g., SPO1A,TNX-355, 9) a gp120 inhibitor, e.g., BMS-488043 and BlockAide/CR, 10) aG6PD and NADH-oxidase inhibitor, e.g., immunitin, 11) a CCR5 inhibitor,e.g., aplaviroc, vicriviroc, INCB9471, PRO-140, INCB15050, PF-232798,CCR5mAb004, and maraviroc, 12) an interferon, e.g., pegylated rIFN-alpha2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b, IFN alpha-2b XL, rIFN-alpha2a, consensus IFN alpha, infergen, rebif, locteron, AVI-005,PEG-infergen, pegylated IFN-beta, oral interferon alpha, feron,reaferon, intermax alpha, r-IFN-beta, infergen+actimmune, IFN-omega withDUROS, and albuferon, 13) ribavirin analogs, e.g., rebetol, copegus,levovirin, VX-497, and viramidine (taribavirin), 14) NS5a inhibitors,e.g., A-831, A-689, and BMS-790052, 15) NS5b polymerase inhibitors,e.g., NM-283, valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433, andXTL-2125, 16) NS3 protease inhibitors, e.g., SCH-503034 (SCH-7), VX-950(Telaprevir), ITMN-191, and BILN-2065, 17) alpha-glucosidase 1inhibitors, e.g., MX-3253 (celgosivir) and UT-231B, 18)hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and LB-84451, 19)non-nucleoside inhibitors of HCV, e.g., benzimidazole derivatives,benzo-1,2,4-thiadiazine derivatives, and phenylalanine derivatives, 20)other drugs for treating Hepatitis C, e.g., zadaxin, nitazoxanide(alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702, AVI 4065,bavituximab, oglufanide, PYN-17, KPE02003002, actilon (CPG-10101),KRN-7000, civacir, GI-5005, ANA-975 (isatoribine), XTL-6865, ANA 971,NOV-205, tarvacin, EHC-18, and NIM811, 20) pharmacokinetic enhancers,e.g., BAS-100 and SPI452, 21) RNAse H inhibitors, e.g., ODN-93 andODN-112, 22) other anti-HIV agents, e.g., VGV-1, PA-457 (bevirimat),ampligen, HRG214, cytolin, polymun, VGX-410, KD247, AMZ 0026, CYT 99007,A-221 HIV, BAY 50-4798, MDX010 (iplimumab), PBS119, ALG889, andPA-1050040.

It is contemplated that the second therapeutic agent will beadministered in a manner that is known in the art and the dosage may beselected by someone of skill in the art. For example, the second agentmay be administered in a dose from about 0.01 milligrams to about 2grams per day.

Metabolites of the Compounds

Also falling within the scope of this disclosure are the in vivometabolic products of the compounds described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,esterification and the like of the administered compound, primarily dueto enzymatic processes. Accordingly, the disclosure includes compoundsproduced by a process comprising contacting a compound of thisdisclosure with a mammal for a period of time sufficient to yield ametabolic product thereof. Such products typically are identified bypreparing a radiolabelled (e.g., C¹⁴ or H³) compound of the disclosure,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, orto man, allowing sufficient time for metabolism to occur (typicallyabout 30 seconds to 30 hours) and isolating its conversion products fromthe urine, blood or other biological samples. These products are easilyisolated since they are labeled (others are isolated by the use ofantibodies capable of binding epitopes surviving in the metabolite). Themetabolite structures are determined in conventional fashion, e.g., byMS or NMR analysis. In general, analysis of metabolites is done in thesame way as conventional drug metabolism studies well-known to thoseskilled in the art. The conversion products, so long as they are nototherwise found in vivo, are useful in diagnostic assays for therapeuticdosing of the compounds of the disclosure even if they possess no HCVinhibitory activity of their own.

Methods for determining stability of compounds in surrogategastrointestinal secretions are known.

Exemplary Methods of Making the Compounds

The disclosure also relates to methods of making the compositions of thedisclosure. The compositions are prepared by any of the applicabletechniques of organic synthesis. Many such techniques are well known inthe art. However, many of the known techniques are elaborated inCompendium of Organic Synthetic Methods (John Wiley & Sons, New York),Vol. 1, Ian T. Harrison and Shuyen Harrison, 1971; Vol. 2, Ian T.Harrison and Shuyen Harrison, 1974; Vol. 3, Louis S. Hegedus and LeroyWade, 1977; Vol. 4, Leroy G. Wade, Jr., 1980; Vol. 5, Leroy G. Wade,Jr., 1984; and Vol. 6, Michael B. Smith; as well as March, J., AdvancedOrganic Chemistry, Third Edition, (John Wiley & Sons, New York, 1985),Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency inModern Organic Chemistry. In 9 Volumes, Barry M. Trost, Editor-in-Chief(Pergamon Press, New York, 1993 printing). Other methods suitable forpreparing compounds of the disclosure are described in InternationalPatent Application Publication Number WO 2006/020276.

A number of exemplary methods for the preparation of the compositions ofthe disclosure are provided in the schemes and examples below. Thesemethods are intended to illustrate the nature of such preparations andare not intended to limit the scope of applicable methods.

Generally, the reaction conditions such as temperature, reaction time,solvents, work-up procedures, and the like, will be those common in theart for the particular reaction to be performed. The cited referencematerial, together with material cited therein, contains detaileddescriptions of such conditions. Typically the temperatures will be−100° C. to 200° C., solvents will be aprotic or protic, and reactiontimes will be 10 seconds to 10 days. Work-up typically consists ofquenching any unreacted reagents followed by partition between awater/organic layer system (extraction) and separating the layercontaining the product.

Oxidation and reduction reactions are typically carried out attemperatures near room temperature (about 20° C.), although for metalhydride reductions frequently the temperature is reduced to 0° C. to−100° C., solvents are typically aprotic for reductions and may beeither protic or aprotic for oxidations. Reaction times are adjusted toachieve desired conversions.

Condensation reactions are typically carried out at temperatures nearroom temperature, although for non-equilibrating, kinetically controlledcondensations reduced temperatures (0° C. to −100° C.) are also common.Solvents can be either protic (common in equilibrating reactions) oraprotic (common in kinetically controlled reactions).

Standard synthetic techniques such as azeotropic removal of reactionby-products and use of anhydrous reaction conditions (e.g., inert gasenvironments) are common in the art and will be applied when applicable.

The terms “treated”, “treating”, “treatment”, and the like, when used inconnection with a chemical synthetic operation, mean contacting, mixing,reacting, allowing to react, bringing into contact, and other termscommon in the art for indicating that one or more chemical entities istreated in such a manner as to convert it to one or more other chemicalentities. This means that “treating compound one with compound two” issynonymous with “allowing compound one to react with compound two”,“contacting compound one with compound two”, “reacting compound one withcompound two”, and other expressions common in the art of organicsynthesis for reasonably indicating that compound one was “treated”,“reacted”, “allowed to react”, etc., with compound two. For example,treating indicates the reasonable and usual manner in which organicchemicals are allowed to react. Normal concentrations (0.01M to 10M,typically 0.1M to 1M), temperatures (−100° C. to 250° C., typically −78°C. to 150° C., more typically −78° C. to 100° C., still more typically0° C. to 100° C.), reaction vessels (typically glass, plastic, metal),solvents, pressures, atmospheres (typically air for oxygen and waterinsensitive reactions or nitrogen or argon for oxygen or watersensitive), etc., are intended unless otherwise indicated. The knowledgeof similar reactions known in the art of organic synthesis is used inselecting the conditions and apparatus for “treating” in a givenprocess. In particular, one of ordinary skill in the art of organicsynthesis selects conditions and apparatus reasonably expected tosuccessfully carry out the chemical reactions of the described processesbased on the knowledge in the art.

Modifications of each of the exemplary schemes and in the Examples(hereafter “exemplary schemes”) leads to various analogs of the specificexemplary materials produced. The above-cited citations describingsuitable methods of organic synthesis are applicable to suchmodifications.

In each of the exemplary schemes it may be advantageous to separatereaction products from one another and/or from starting materials. Thedesired products of each step or series of steps is separated and/orpurified (hereinafter separated) to the desired degree of homogeneity bythe techniques common in the art. Typically such separations involvemultiphase extraction, crystallization from a solvent or solventmixture, distillation, sublimation, or chromatography. Chromatographycan involve any number of methods including, for example: reverse-phaseand normal phase; size exclusion; ion exchange; high, medium, and lowpressure liquid chromatography methods and apparatus; small scaleanalytical; simulated moving bed (SMB) and preparative thin or thicklayer chromatography, as well as techniques of small scale thin layerand flash chromatography.

Another class of separation methods involves treatment of a mixture witha reagent selected to bind to or render otherwise separable a desiredproduct, unreacted starting material, reaction by product, or the like.Such reagents include adsorbents or absorbents such as activated carbon,molecular sieves, ion exchange media, or the like. Alternatively, thereagents can be acids in the case of a basic material, bases in the caseof an acidic material, binding reagents such as antibodies, bindingproteins, selective chelators such as crown ethers, liquid/liquid ionextraction reagents (LIX), or the like.

Selection of appropriate methods of separation depends on the nature ofthe materials involved. For example, boiling point, and molecular weightin distillation and sublimation, presence or absence of polar functionalgroups in chromatography, stability of materials in acidic and basicmedia in multiphase extraction, and the like. One skilled in the artwill apply techniques most likely to achieve the desired separation.

A single stereoisomer, e.g., an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as formation of diastereomers using optically activeresolving agents (Stereochemistry of Carbon Compounds, (1962) by E. L.Eliel, McGraw Hill; Lochmuller, C. H., (1975) J. Chromatogr., 113, 3)283-302). Racemic mixtures of chiral compounds of the disclosure can beseparated and isolated by any suitable method, including: (1) formationof ionic, diastereomeric salts with chiral compounds and separation byfractional crystallization or other methods, (2) formation ofdiastereomeric compounds with chiral derivatizing reagents, separationof the diastereomers, and conversion to the pure stereoisomers, and (3)separation of the substantially pure or enriched stereoisomers directlyunder chiral conditions.

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as brucine, quinine, ephedrine,strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like withasymmetric compounds bearing acidic functionality, such as carboxylicacid and sulfonic acid. The diastereomeric salts may be induced toseparate by fractional crystallization or ionic chromatography. Forseparation of the optical isomers of amino compounds, addition of chiralcarboxylic or sulfonic acids, such as camphorsulfonic acid, tartaricacid, mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(Eliel, E. and Wilen, S. (1994) Stereochemistry of Organic Compounds,John Wiley & Sons, Inc., p. 322). Diastereomeric compounds can be formedby reacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as menthyl derivatives, followed byseparation of the diastereomers and hydrolysis to yield the free,enantiomerically enriched substrate. A method of determining opticalpurity involves making chiral esters, such as a menthyl ester, e.g., (−)menthyl chloroformate in the presence of base, or Mosher ester,α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob III. (1982) J. Org.Chem. 47:4165), of the racemic mixture, and analyzing the NMR spectrumfor the presence of the two atropisomeric diastereomers. Stablediastereomers of atropisomeric compounds can be separated and isolatedby normal- and reverse-phase chromatography following methods forseparation of atropisomeric naphthyl-isoquinolines (Hoye, T., WO96/15111). By method (3), a racemic mixture of two enantiomers can beseparated by chromatography using a chiral stationary phase (ChiralLiquid Chromatography (1989) W. J. Lough, Ed. Chapman and Hall, NewYork; Okamoto, (1990) J. of Chromatogr. 513:375-378). Enriched orpurified enantiomers can be distinguished by methods used to distinguishother chiral molecules with asymmetric carbon atoms, such as opticalrotation and circular dichroism.

Schemes and Examples

General aspects of these exemplary methods are described below and inthe Examples. Each of the products of the following processes isoptionally separated, isolated, and/or purified prior to its use insubsequent processes.

A number of exemplary methods for the preparation of compounds of thedisclosure are provided herein, for example, in the Examples below.These methods are intended to illustrate the nature of such preparationsand are not intended to limit the scope of applicable methods. Certaincompounds of the disclosure can be used as intermediates for thepreparation of other compounds of the disclosure. In the exemplarymethods described herein, the fragment E-V— can also be written as R9-.PG represents a protecting group common for the given functional groupthat it is attached. The installation and removal of the protectinggroup can be accomplished using standard techniques, such as thosedescribed in Wuts, P. G. M., Greene, T. Protective Groups in OrganicSynthesis, 4th ed.; John Wiley & Sons, Inc.: Hoboken, N.J., 2007.

Scheme 1 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V-Emolecule of the disclosure wherein, for illustrative purposes, E ismethoxycarbonylamino. The treatment of either 1a or 1c with one or twoequivalents respectively of methyl chloroformate under basic conditions(e.g. sodium hydroxide) provides the molecule 1b or 1d.

Scheme 2 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V-Emolecule of the disclosure wherein, for illustrative purposes, P ispyrrolidine. Coupling of amine 2a with acid 2b is accomplished using apeptide coupling reagent (e.g. HATU) to afford 2c. Alternatively, amine2d is coupled with two equivalents of 2b under similar conditions toprovide 2e.

Scheme 6 shows a general synthesis of an R¹—V—C(═O)—P—R² intermediatewherein, for illustrative purposes, P is pyrrolidine, R¹ is a genericgroup that is depicted as either -E or a amino protecting group, and R²is a generic group that is depicted as —W—P—C(═O)—V-E,—W—P—C(═O)—V—NH-PG, —W—P—NH-PG, or —W—NH-PG. Coupling of amine 6a (or6d, 6h, 6k) with acid 6b or 6e is accomplished using a peptide couplingreagent (e.g. HATU) to afford 6c (or 6f, 6g, 6i, 6j, 6l, 6m)respectively.

Scheme 7 shows a general synthesis of an E-V—C(═O)—R¹ intermediatewherein, for illustrative purposes, E is methoxycarbonylamino and R¹ isa generic group that is depicted as either —P—W—P—C(═O)—V—NH-PG,—P—W—P—PG, —P—W—PG, —P—PG, or —O—PG. Treatment of 7a (or 7c, 7e, 7g, 7i)with methyl chloroformate under basic conditions (e.g. sodium hydroxide)provides the molecule 7b (or 7d, 7f, 7h, 7j).

Scheme 9 shows a general synthesis of an R¹—P—R² intermediate wherein,for illustrative purposes, R¹ is —C(═O)—V-E or a protecting group and R²is a substituted benzimidazole. The formation of the benzimidazole isaccomplished by coupling the acid 9b or 9e with an arylamine 9a, using apeptide coupling reagent such as HATU, to afford 9c or 9f. Cyclizationof the amide in the presence of an acid (such as acetic acid) affordsthe benzimidazole containing molecule 9d or 9g.

The formation of multiple benzimidazoles is performed in the samemanner, starting with a bis-diamine to provide the correspondingbis-benzimidazole.

Scheme 20 shows a general synthesis of an R¹—P—W—P—R² intermediate ofthe disclosure wherein, for illustrative purposes, R¹ and R² areindependent protecting groups and W is a two aromatic ring unitconstructed via a transition metal mediated cyclization. Alkylation ofphenol 20b with an alkyl bromide, such as 20a, provides the ether 20c.Cyclization of the aromatic rings in the presence of a palladiumcatalyst provides the compound 20d. Treatment of 20d with CuBr₂ providesthe α-haloketone 20e, which provides 20f upon addition of an acid underbasic conditions (e.g. Et3N). Reaction of 20f with an amine or aminesalt (e.g. ammonium acetate) affords the imidazole containing molecule20g. Oxidation of 20g, 20i, or 20l can be accomplished by heating in thepresence of MnO₂ to provide 20h, 20j, or 20m, respectively. Conversionof 20g or 20h with a palladium catalyst, such as Pd₂dba₃ and X-Phos, anda boron source such as bis(pinacolato)diboron provides the boronic ester20i or 20j. The boronic ester is coupled with an appropriate couplingpartner (e.g. 20k) using a palladium catalyst, such as Pd(PPh₃)₄ orPdCl₂(dppf), to afford 20l or 20m. For each transition metal mediatedcross-coupling reaction, the roles of the nucleophile and electrophilecan be reversed to provide the same coupling product. Other transitionmetal mediated cross couplings that enable the construction of W, butemploy alternative coupling partners and reagents, include, but are notlimited to, the Negishi, Kumada, Stille, and Ullman couplings. For thepreparation of alternate two aromatic ring containing W groups, thisgeneral scheme can be applied through the appropriate choice of thestarting reagents.

Scheme 21 shows a general synthesis of an R¹—P—W—P—R² intermediate ofthe disclosure wherein, for illustrative purposes, R¹ and R² areindependent protecting groups and W is a two aromatic ring unitconstructed via a transition metal mediated cyclization. Treatment of20d with an activated vinyl reagent (e.g. potassiumvinyltrifluoroborate) in the presence of a palladium catalyst (e.g.palladium acetate and S-Phos) provides the vinyl compound 21a.Conversion to the corresponding α-halo ketone can be accomplished bybromination with N-bromosuccinimide, followed by oxidation with MnO₂.Displacement of the α-halo ketone proceeds by the addition of an acidunder basic conditions (e.g. Et₃N). Bromination of 21c proceeds upontreatment with pyridinium tribromide, and is followed by the addition ofa second acid under basic conditions to provide the diester 21e.Reaction of 21e with an amine or amine salt (e.g. ammonium acetate)affords the imidazole containing molecule 21f. Oxidation of 21f can beaccomplished in the presence of MnO₂ to provide 21g.

Scheme 22 shows a general synthesis of an E-V—C(═O)—P—W—P—R intermediateof the disclosure wherein, for illustrative purposes, R is a protectinggroup and W is a two aromatic ring unit. Displacement of the α-haloketone 21b proceeds by the addition of an acid under basic conditions(e.g. Et₃N). Bromination of 22a proceeds upon treatment with pyridiniumtribromide, and is followed by the addition of a second acid under basicconditions to provide the diester 22c. Reaction of 22c with an amine oramine salt (e.g. ammonium acetate) affords the imidazole containingmolecule 22d. Oxidation of 22d can be accomplished in the presence ofMnO₂ to provide 22e.

Scheme 23 shows a general synthesis of an E-V—C(═O)—P—W—P—R intermediateof the disclosure wherein, for illustrative purposes, R is a protectinggroup and W is a two aromatic ring unit. Displacement of the α-haloketone 21d proceeds by the addition of an acid under basic conditions(e.g. Et₃N). Reaction of 23a with an amine or amine salt (e.g. ammoniumacetate) affords the imidazole containing molecule 23b. Oxidation of 23bcan be accomplished in the presence of MnO₂ to provide 23c.

Scheme 25 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V-Emolecule of the disclosure wherein, for illustrative purposes, E isethylcarbonylamino. The treatment of either 25a or 25c with one or twoequivalents respectively of propionyl chloride under basic conditions(e.g. sodium hydroxide) provides the molecule 25b or 25d.

Scheme 26 shows a general synthesis of an E-V—C(═O)—P—R and an R¹—P—Rmolecule of the disclosure wherein, for illustrative purposes R is ahaloimidazole. Treatment of the aldehyde 26a with glyoxal, in thepresence of ammonium hydroxide provides the imidazole 26b. Treatmentwith either N-bromosuccinamide or iodine provides the correspondinghaloimidazole 26c and 26d respectively. Separation from thecorresponding bis-halogenated compound can be accomplished bypreparative HPLC chromatography. The conversion of the bis-haloimidazoleto the mono-haloimidazole can also be accomplished upon heating in thepresence of sodium sulfite. Further functionalization of the P group canbe accomplished upon removal of the protecting group and coupling withan appropriate acid (E-V—C(═O)—OH).

Scheme 27 shows an alternate general synthesis of an R¹—P—W—P—R²intermediate of the invention wherein, for illustrative purposes, R¹ andR² are independent protecting groups and W is a two aromatic ring unitconstructed via a transition metal mediated cyclization. Bromination of21b with a brominating agent (i.e. pyridinium tribromide) provides thedibromide 27a. Displacement of the primary bromide then proceeds by theaddition of an acid under basic conditions (e.g. K₂CO₃) to provide 21d.Conversion to 21f or 21g can be accomplished following methods describedin Scheme 21.

Scheme 28 shows an alternate general synthesis of an E-V—C(═O)—P—W—P—Rintermediate of the invention wherein, for illustrative purposes, R is aprotecting group and W is a two aromatic ring unit. Bromination of 21bwith a brominating agent (i.e. pyridinium tribromide) provides thedibromide 27a. Displacement of the primary bromide then proceeds by theaddition of an acid under basic conditions (e.g. K₂CO₃) to provide 22d.Conversion to 22d or 22e can be accomplished following methods describedin Scheme 22.

Specific Embodiments

In one embodiment the disclosure provides a compound which has formula:

wherein the imidazole ring shown in formula A1, A2, A3, and A4 isoptionally substituted with one or more groups independently selectedfrom halo, haloalkyl, cyano, or alkyl;

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment the disclosure provides a compound which has formula:

wherein the imidazole ring shown in formula A2 and A4 is optionallysubstituted with one or more groups independently selected from halo,haloalkyl, cyano, or alkyl;

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment at least one of E^(1a) and E^(1b) is—N(H)(alkoxycarbonyl).

In one embodiment at least one of E^(1a) and E^(1b) is —N(H)C(═O)OMe.

In one embodiment both of E^(1a) and E^(1b) are —N(H)C(═O)OMe.

In one embodiment at least one of E^(1a) and E^(1b) is—N(H)(cycloalkylcarbonyl) or —N(H)(cycloalkyloxycarbonyl).

In one embodiment at least one of E^(1a) and E^(1b) iscyclopropylcarbonylamino, cyclobutylcarbonylamino,cyclopropyloxycarbonylamino or cyclobutyloxycarbonylamino.

In one embodiment E^(1a) and E^(1b) are each independently selected fromcyclopropylcarbonylamino, cyclobutylcarbonylamino,cyclopropyloxycarbonylamino or methoxycarbonylamino.

In one embodiment at least one of V^(1a) and V^(1b) is:

In one embodiment E^(1a)-V^(1a) taken together are R^(9a) or whereinE^(1b)-V^(1b) taken together are R^(9b).

In one embodiment at least one of P^(1a) and P^(1b) is selected from:

In one embodiment P^(1a) and P^(1b) are each independently selectedfrom:

In one embodiment one of P^(1a) and P^(1b) is:

and the other of P^(1a) and P^(1b) is:

In one embodiment one of P^(1a) and P^(1b) is:

and the other of P^(1a) and P^(1b) is:

In one embodiment at least one of P^(1a) and P^(1b) is:

In one embodiment at least one of P^(1a) and P^(1b) is:

In one embodiment at least one of —V^(1a)—C(═O)—P^(1a) and—P^(1b)—C(═O)—V^(1b)— is:

In one embodiment at least one of —V^(1a)—C(═O)—P^(1a) and—P^(1b)—C(═O)—V^(1b)— is:

In one embodiment both of —V^(1a)—C(═O)—P^(1a) and —P^(1b)—C(═O)—V^(1b)—are independently selected from:

In one embodiment one of —V^(1a)—C(═O)—P^(1a) and —P^(1b)—C(═)—V^(1b)—is:

and the other of —V^(1a)—C(═O)—P^(1a) and —P^(1b)—C(═O)—V^(1b)— is:

In one embodiment the disclosure provides a compound of formula:

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment the disclosure provides a compound of formula:

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment the disclosure provides a compound of formula:

or a pharmaceutically acceptable salt or prodrug thereof.

The disclosure will now be illustrated by the following non-limitingExamples. The following abbreviations are used throughout thespecification, including the Examples.

(aq) Aqueous (g) Gas (s) Solid ° C. Degree Celsius Ac Acetate ACNAcetonitrile apprx Approximate Bis- Bis(pinacolato)diboronpinB/(Bpin)₂/(pinB)₂ BOC/Boc tert-Butoxycarbonyl calc'd Calculated CC₅₀50% Cytotoxicity concentration COMU 1-[(1-(Cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethyl- aminomorpholino)] uroniumhexafluorophosphate d Doublet dba dibenzalacetone DCM Dichloromethane ddDoublet of doublets ddd Doublet of doublet of doublets DIPEA/DIEAN,N-Diisopropylethylamine DMA N,N-Dimethylacetamide DMAP4-Dimethylaminopyridine DME Dimethoxyethane DMEM Eagle's minimalessential medium DMF Dimethylformamide DMSO/dmso Dimethylsulfoxide dppf1,1′-bis(diphenylphosphanyl) ferrocene dt Doublet of triplets EC₅₀ Halfmaximal effective concentration ESI Electrospray ionization Et Ethylext. External FBS Fetal bovine serum g Gram HATU2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3- tetramethyl uroniumhexafluorophosphate Methanaminium HPLC High performance liquidchromatography hr/h Hour Hz Hertz J Coupling constant LCMS Liquidchromatography mass spectrometry M Molar m Multiplet m/z Mass to chargeM+ Mass peak Me Methyl mg Milligram MHz Megahertz min Minute mLMilliliter mmol Millimole Moc Methoxycarbonyl MS Mass spectrometry MTBEMethyl tert-butyl ether N Normal NADPH Nicotinamide adenine dinucleotidephosphate NBS N-Bromosuccinimide NMM N-Methylmorpholine NMR Nuclearmagnetic resonance o/n Over night Papp Apparent permeability PBSPhosphate buffer system Pd/C Palladium on carbon Ph Phenyl Phg/PhGlyPhenyl glycine Piv Pivalate Pro Proline pyr Pyridine q Quartet qdQuartet of doublets quant Quantitative quint Quintet rt/RT Roomtemperature s Singlet SPhos 2-Dicyclohexylphosphino-2′,6′-dimethoxybiphenyl t Triplet t-Bu tert-Butyl TEMPO(2,2,6,6-Tetramethyl-piperidin-1-yl)oxyl Tf TrifluoromethanesulfonateTFA Trifluoroacetic acid THF Tetrahydrofuran Thr Threonine TLC Thinlayer chromatography tol. Toluene UV Ultraviolet Val Valine w/v Weightto volume w/w Weight to weight X-Phos/XPOS/Xphos2-Dicyclohexylphosphino-2′,4′,6′- triisopropylbiphenyl δ Chemical shiftμg Microgram μL Microliter

EXAMPLES Example LQ

7-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one

To a stirred solution of 7-hydroxy-1-tetralone (13.9 g, 85.7 mmol) and1-bromo-2-(bromomethyl)-4-chlorobenzene (25.6 g, 90.0 mmol) indimethylformamide (850 mL) was added potassium carbonate (24 g, 172mmol). The reaction was stirred under argon for 18 hours then dilutedwith ethyl acetate (1 L). The organics were washed three times withwater and once with brine. The organic layer was then dried withmagnesium sulfate, filtered and concentrated. To the resulting oil wasadded methanol (500 mL) and the suspension was agitated for thirtyminutes. 7-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one(27.8 g, 89% yield) was isolated by filtration.

3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

To a 1 L flask containing palladium(II) pivalate (1.18 g, 3.8 mmol),tri(4-fluorophenyl)phosphine (1.20 g, 3.8 mmol), pivalic acid (2.33 g,22.8 mmol) and potassium carbonate (31.8 g, 228 mmol) was added asolution of7-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one (27.8 g,76.2 mmol) in dimethylacetamide (380 mL). The flask was evacuated andbackfilled with argon 5 times and then stirred under argon at 60° C. for24 hours. The reaction was cooled to room temperature and diluted withMTBE and water. The resulting biphasic mixture was stirred for 3 hoursand filtered through Celite, rinsing with MTBE. The organic layer of thefiltrate was separated and then washed twice with water and once withbrine. The organics were then dried with magnesium sulfate, filtered,concentrated and purified by flash column chromatography (Hexanes/DCM)to yield 3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (14.4g, 67% yield) as an off-white solid.

9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

To a mixture of 3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(14.8 g, 52 mmol) in chloroform (50 mL) and ethyl acetate (50 mL) wasadded copper(II) bromide (24.3 g, 104 mmol). The reaction was heated to80° C. for 2 hours and then cooled to room temperature. The mixture wasdiluted with dichloromethane and washed twice with a 5:1 solution ofsaturated aqueous ammonium chloride and aqueous ammonium hydroxide(˜38%), and washed once with water. The organic layer was dried withmagnesium sulfate, filtered and concentrated to yield9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (18.5g, >95% yield) with >95% purity.

Note: This reaction is not always this clean. Sometimes there isover-bromination and sometimes there is significant starting material.These impurities can be removed by flash column chromatography.

Tert-Butyl2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

To a solution of (1R)-2-(tert-butoxycarbonyl)cyclopentanecarboxylic acid(10.17 g, 47.25 mmol) and9-bromo-3-chloro-10,11-dihydro-6H-naphtho[2,3-c]chromen-8(9H)-one (5.7mg, 15.7 mmol) in acetonitrile (50 mL) was added diisopropylethylamine(11.11 mL, 64 mmol). The reaction was stirred at 50° C. for 4 hours andwas then diluted with ethyl acetate. The organics were washed with waterand brine, dried (MgSO₄) and concentrated. The resulting crude residuewas purified by flash chromatography to yield (2S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-naphtho[c,g]chromen-9-yl)pyrrolidine-1,2-dicarboxylate (4.52 g, 58%). To a solution of(2S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-6H-naphtho[2,3-c]chromen-9-yl)pyrrolidine-1,2-dicarboxylate (3.27 mg, 6.56 mmol) in a mixture oftoluene (11 mL) and 2-methoxyethanol (0.7 mL) was added ammonium acetate(5.06 g, 65.6 mmol). The reaction mixture was heated to 110° C. for 3hours, cooled to room temperature and diluted with ethyl acetate. Theorganics were washed with water and brine, dried (Na₂SO₄), andconcentrated. The crude residue was purified by flash chromatography toyield tert-butyl2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.95 g, 61%). LCMS-ESI⁺: calculated for C27H28ClN₃O3₄ ₂: 477.98;observed [M+1]⁺: 478.47

Tert-Butyl2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

To a solution of tert-butyl2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.9 g, 3.96 mmol) in dichloromethane (35 mL) was added manganese(IV)oxide (17 g, 198 mmol). The reaction mixture was stirred at roomtemperature for 18 hours, diluted with ethyl acetate. The organics werewashed with water and brine, dried (Na₂SO₄), and concentrated. The cruderesidue was purified by flash chromatography to yield tert-butyl2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.52 g, 81%). LCMS-ESI⁺: calculated for C27H26ClN3O3: 475.9; observed[M+1]⁺: 476.45.

Tert-Butyl2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate

A degassed mixture of tert-butyl2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.52 g, 3.17 mmol), bis(pinacolato)diboron (1.21 g, 4.75 mmol),potassium acetate (934 mg, 9.52 mmol),tris(dibenzylideneacetone)palladium (116 mg, 0.13 mmol) and2-dicyclohexylphosphino-2′, 4′, 6′-tri-i-propyl-1, 1′-biphenyl (121 mg,0.08 mmol) in 1,4-dioxane (16 mL) was heated to 90° C. for 1.5 hours,cooled to room temperature and diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield tert-butyl2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(1.7 g, 94%)

Tert-Butyl2-[9-(2-{1-[N-(methoxycarbonyl)valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate

To a solution of methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(1.48 g, 3.97 mmol), tert-butyl2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(1.88 g, 1.48 mmol), tetrakis(triphenyl phosphine)palladium(0) (191 mg,0.16 mmol) and dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (242 mg, 0.33 mmol) in a mixture of1,2-dimethoxyethane (37.0 mL) and dimethylformamide (6 mL) was added asolution of potassium carbonate (2M in water, 5 mL, 9.93 mmol). Theresulting mixture was degassed and then heated to 85° C. under argon for18 hours. After cooling to room temperature, the reaction was dilutedwith ethyl acetate. The organics were washed with water and brine, dried(Na₂SO₄), and concentrated. The crude residue was purified by flashchromatography to yield tert-butyl2-[9-(2-{1-[N-(methoxycarbonyl)valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(1.45 mg, 59%). LCMS-ESI⁺: calculated for C41H47N7O6₇₃ 733.86; observed[M+1]⁺: 734.87.

[1-(2-{5-[2-(1-{[(Methoxycarbonyl)amino](phenyl)acetyl}pyrrolidin-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl]-1H-imidazol-2-yl}pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl]carbamicAcid

A solution of tert-butyl2-[9-(2-{1-[N-(methoxycarbonyl)valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(462 mg, 0.63 mmol), ethanol (6 mL) and concentrated HCl (2 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (172 mg, 0.82 mmol) andCOMU (311 mg, 0.73 mmol) in DMF (6 mL). To the resulting solution wasadded diisopropylethylamine (330 μL, 1.89 mmol). After stirring for 18hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 45% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give[1-(2-{5-[2-(1-{[(methoxycarbonyl)amino](phenyl)acetyl}pyrrolidin-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl]-1H-imidazol-2-yl}pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl]carbamicacid (231 mg, 45%). LCMS-ESI⁺: calculated for C46H48N8O7₈: 824.92;observed [M+1]⁺: 826.00.

Example LR

(2S,4S)-1-tert-butyl 2,4-dimethyl pyrrolidine-1,2,4-tricarboxylate

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-cyanopyrrolidine-1,2-dicarboxylate (9.0 g, 35.4 mmol) in MeOH (196 mL)was added HCl (4M in 1,4-dioxane, 100 mL, 403 mmol). The solution wasstirred at room temperature for 16h and concentrated in vacuo. The crudeintermediate was dissolved in EtOAc (180 mL) and basified with aqueousbicarbonate (sat.). Di-tert-butyl dicarbonate (8.5 g, 38.9 mmol) wasadded and the biphasic solution was stirred at room temperature for 12h.The layers were then separated and the aqueous layer was back extractedwith EtOAc. The combined organic layers were washed with brine, driedover Na₂SO₄, and concentrated. The crude oil was purified by silica gelchromatography (15% to 40% to 100% EtOAc/Hexanes) to provide(2S,4S)-1-tert-butyl 2,4-dimethyl pyrrolidine-1,2,4-tricarboxylate (9.56g, 94%).

(3S,5S)-1-(tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicAcid

To a solution of (2S,4S)-1-tert-butyl 2,4-dimethylpyrrolidine-1,2,4-tricarboxylate (9.56 g, 33.3 mmol) in THF (70 mL) at0° C. (external temperature, ice bath) was added NaOH (1N aqueous, 33mL, 33.3 mmol) dropwise over 15 min. The solution was stirred at 0° C.for 5h before acidification with HCl (1N). The solution was extractedwith EtOAc (3×). The combined organic layers were dried over Na₂SO₄ andconcentrated. The crude oil was purified by silica gel chromatography(2% to 5% to 10% MeOH/CH₂Cl₂) to provide(3S,5S)-1-(tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicacid (6.38g, 70%).

(2S,4S)-1-tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate

To a solution of(3S,5S)-1-(tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicacid (6.38 g, 23.3 mmol) in THF (116 mL) at 0° C. (external temperature,ice bath) was added Et₃N (4.9 mL, 35.0 mmol) and ethyl chloroformate(2.7 mL, 28.0 mmol). The resulting solution was stirred at 0° C. for 45min, during which time a white precipitate forms. The reaction mixturewas filtered through celite and concentrated.

The crude intermediate was dissolved in THF (59 mL) and cooled to 0° C.(external temperature, ice bath). NaBH₄ (4.41 g, 116.7 mmol) in H₂O (59mL) was slowly added and the resulting solution was stirred at 0° C. for2 h. The reaction mixture was diluted with EtOAc and washed with H₂O.The aqueous layer was backextracted with EtOAc. The combined organiclayers were dried over Na₂SO₄ and concentrated. The crude oil waspurified by silica gel chromatography (42% to 69% to 100% EtOAc/Hexanes)to provide (2S,4S)-1-tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate (3.63 g, 60%).

(2S,4S)-1-tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate (2.57 g, 9.9 mmol) inCH₂Cl₂ (50 mL) was added AgOTf (4.07 g, 15.8 mmol) and2,6-di-tert-butylpyridine (4.4 mL, 19.8 mmol). The reaction mixture wascooled to 0° C. (external temperature, ice bath) and Met (0.98 mL, 15.8mmol) was slowly added. The resulting slurry was stirred at 0° C. for1.5 h and at room temperature for 1.5 h. The slurry was diluted withCH₂Cl₂ and filtered through celite. The filtrate was concentrated todryness, dissolved in Et₂O, and washed with HCl (1N) and brine. Theaqueous layers were backextracted with Et₂O and the combined organiclayers were dried over Na₂SO₄ and concentrated. The crude oil waspurified by silica gel chromatography (10% to 75% to 100% EtOAc/Hexanes)to provide (2S,4S)-1-tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (2.11 g, 78%). ¹H-NMR:400 MHz, (CDCl₃) δ: (mixture of rotamers, major reported) 4.20 (t, 1H),3.71 (s, 3H), 3.67 (m, 1H), 3.34 (m, 2H), 3.30 (s, 3H), 3.16 (t, 1H),2.43 (m, 2H), 1.74 (m, 1H), 1.38 (s, 9H).

(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (2.11 g, 7.7 mmol) in amixture of THF (38 mL) and MeOH (15 mL) was added LiOH (2.5 M aqueous,15 mL, 38.6 mmol). The resulting solution was stirred at roomtemperature for 2h, and acidified with aqueous HCl (1N). The desiredproduct was extracted with CH₂Cl₂ (4×). The combined organic layers weredried over Na₂SO₄ and concentrated to provide(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (2.0 g, 99%). ¹H-NMR: 400 MHz, (CDCl₃) δ: (mixture of rotamers,major reported) 4.33 (t, 1H), 3.65 (m, 1H), 3.35 (m, 2H), 3.32 (s, 3H),3.16 (t, 1H), 2.45 (m, 2H), 2.12 (m, 1H), 1.46 (s, 9H).

Example LR-1

(2S,4S)-tert-butyl-2-(9-chloro-4,5-dihydro-5H-naphtho[2,3-c]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of((S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (5.9 g, 23.1 mmol) and9-bromo-3-chloro-10,11-dihydro-5H-naphtho[c,g]chromen-8(9H)-one (5.6 mg,15.4 mmol) in acetonitrile (60 mL) was added diisopropylethylamine (5.35mL, 30.8 mmol). The reaction was stirred at 50° C. for 18 hours and wasthen diluted with ethyl acetate. The organics were washed with water andbrine, dried (MgSO₄) and concentrated. The resulting crude residue waspurified by flash chromatography to yield(2S)-1-tert-butyl-2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-6H-naphtho[2,3-c]chromen-9-yl)-4(methoxymethyl)pyrrolidine-1,2-dicarboxylate (5.12 g, 61%). To a solution of(2S)-1-tert-butyl-2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-6H-naphtho[2,3-c]chromen-9-yl)-4(methoxymethyl)pyrrolidine-1,2-dicarboxylate(5.11 mg, 9.42 mmol) in a mixture of toluene (94 mL) and 2-methoxyethanol (0.1 mL) was added ammonium acetate (23.5 g, 304 mmol). Thereaction mixture was heated to 110° C. for 18 hours, cooled to roomtemperature and diluted with ethyl acetate. The organics were washedwith water and brine, dried (Na₂SO₄), and concentrated. The cruderesidue was purified by flash chromatography to yield (2S,4R)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(1.05g, 21%) and(2S,4S)-tert-butyl-2-(9-chloro-4,5-dihydro-6H-naphtho[2,3-c]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(2.0 g, 41%). LCMS-ESI⁺: calculated for C₂₉H₃₂ClN₃O₄ ₂: 522.0; observed[M+1]⁺: 522.2.

(2S,4S)-tert-butyl-2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of(2S,4S)-tert-butyl-2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(1.99 g, 3.82 mmol) in dichloromethane (30 mL) was added manganese(IV)oxide (10 g, 115 mmol). The reaction mixture was stirred at roomtemperature for 18 hours, diluted with ethyl acetate. The organics werewashed with water and brine, dried (Na₂SO₄), and concentrated. The cruderesidue was purified by flash chromatography to yield(2S,4S)-tert-butyl-2-(9-chloro-6H-naphtho[2,3-c]chromeno[8,9-d]imidazol-2-yl)-4-methoxymethyl)pyrrolidine-1-carboxylate(1.05g, 21%) and(2S,4S)-tert-butyl-2-(9-chloro-4,5-dihydro-6H-naphtho[2,3-c]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(1.64 g, 82%). LCMS-ESI⁺: calculated for C₂₉H₃₀ClN₃O₄₂: 520.02; observed[M+1]⁺: 520.97.

(2S,4S)-tert-butyl-4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)pyrrolidine-1-carboxylate

A degassed mixture of(2S,4S)-tert-butyl-2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (649 mg 1.25 mmol), bis(pinacolato)diboron(635 mg, 2.5 mmol), potassium acetate (368 mg, 3.7 mmol),tris(dibenzylideneacetone)palladium (46 mg, 0.05 mmol) and2-dicyclohexylphosphino-2′, 4′, 6′-tri-i-propyl-1, 1′-biphenyl (60 mg,0.12 mmol) in 1,4-dioxane (7 mL) was heated to 90° C. for 3 hours,cooled to room temperature and diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield(2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)pyrrolidine-1-carboxylate(467 mg, 61%) LCMS-ESI⁺: calculated for C₃₅H₄₂BN₃O₆: 611.54; observed[M+1]⁺: 612.96.

(2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)pyrrolidine-1-carboxylate(467 mg, 0.76 mmol), methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(342 mg, 0.92 mmol), tetrakis(triphenylphosphine) palladium(0) (44 mg,0.04 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (56 mg, 0.07mmol) in a mixture of 1,2-dimethoxyethane (11.0 mL) anddimethylformamide (1.9 mL) was added a solution of potassium carbonate(2M in water, 1.15 mL, 2.29 mmol). The resulting mixture was degassedand then heated to 85° C. under argon for 18 hours. After cooling toroom temperature, the reaction was diluted with ethyl acetate. Theorganics were washed with water and brine, dried (Na₂SO₄), andconcentrated. The crude residue was purified by flash chromatography toyield (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (180mg, 67%). LCMS-ESI⁺: calculated for C₄₃H₅₁₁N₇O₇₃ 777.91; observed[M+1]⁺: 778.84.

methyl{2-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(196 mg, 0.25 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (69 mg, 0.33 mmol) andCOMU (124 mg, 029 mmol) in DMF (4 mL). To the resulting solution wasadded diisopropylethylamine (130 μL, 0.76 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 45% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{2-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(84 mg, 39%). LCMS-ESI⁺: calculated for C₄₂H₅₂N₈O₈: 868.98; observed[M+1]⁺: 870.11

Example LS

methyl{1-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(116 mg, 0.15 mmol), ethanol (5 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (10 mL). This solution was concentrated and tothis material was added a solution of2-methoxycarbonylamino-3-methylbutyric acid (38 mg, 0.22 mmol) and HATU(79 mg, 0.21 mmol) in DMF (1.4 mL). To the resulting solution was addeddiisopropylethylamine (2704, 1.5 mmol). After stirring for 18 hours atroom temperature, the reaction was diluted with ethyl acetate, washedwith water and brine, dried (Na₂SO₄), concentrated and purified bypreparative reverse phase HPLC (Gemini, 15 to 45% ACN/H₂O+0.1% TFA). Theproduct fractions were lyophilized to give methyl{1-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(58 mg, 13%). LCMS-ESI⁺: calculated for C₄₅H₅₄N₈O₈: 834.96; observed[M+1]⁺: 835.70.

Example LT

(2S,4S)-tert-butyl-2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)pyrrolidine-1-carboxylate(557 mg, 0.91 mmol), methyl(S)-1-((1S,3S,5S)-3-(5-bromo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate(350 mg, 0.91 mmol), tetrakis(triphenylphosphine) palladium(0) (53 mg,0.04 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (67 mg, 0.07mmol) in a mixture of 1,2-dimethoxyethane (11.0 mL) anddimethylformamide (1.9 mL) was added a solution of potassium carbonate(2M in water, 1.37 mL, 2.7 mmol). The resulting mixture was degassed andthen heated to 85° C. under argon for 18 hours. After cooling to roomtemperature, the reaction was diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield(2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(271 mg, 38%). LCMS-ESI⁺: calculated for C44H51N7O7. 789.92; observed[M+1]⁺: 790.76.

methyl{2-[2-{9-[2-(2-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(196 mg, 0.25 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (69 mg, 0.33 mmol) andCOMU (124 mg, 0.29 mmol) in DMF (4 mL). To the resulting solution wasadded diisopropylethylamine (130 μL, 0.76 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 45% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{2-[2-{9-[2-(2-{2-[(methoxycarbonyl)aminol-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (84 mg, 39%).LCMS-ESI⁺: calculated for C49H52N8O8: 880.99; observed [M+1]⁺: 882.09

Example LZ

5-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one

To a stirred solution of 5-hydroxy-1-tetralone (2.0 g, 12.3 mmol) and1-bromo-2-(bromomethyl)-4-chlorobenzene (3.6 g, 12.7 mmol) indimethylformamide (125 mL) was added potassium carbonate (3.5 g, 25.1mmol). The reaction was stirred under argon for 1 hour then diluted withethyl acetate (1 L). The organics were washed three times with water andonce with brine. The organic layer was then dried with magnesiumsulfate, filtered and concentrated. To the resulting oil was addedmethanol (100 mL) and the suspension was agitated for thirty minutes.5-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1 (2H)-one (4.25 g,94% yield) was isolated by filtration.

8-chloro-2,3,4,6-tetrahydro-1H-dibenzo[c,h]chromen-1-one

To a flask containing palladium(II) pivalate (68 mg, 0.22 mmol),tri(4-fluorophenyl)phosphine (70 mg, 0.22 mmol), pivalic acid (135 mg,1.3 mmol) and potassium carbonate (1.83 g, 13.1 mmol) was added asolution of5-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one (1.61 g,4.4 mmol) in dimethyacetamide (23 mL). The flask was evacuated andbackfilled with argon 5 times and then stirred under argon at 60° C. for24 hours. The reaction was poured directly onto a silica gel column andpurified by flash column chromatography (hexanes/DCM) to yield8-chloro-2,3,4,6-tetrahydro-1H-dibenzo[c,h]chromen-1-one (1.22 g, 97%yield) as an off-white solid.

2-bromo-8-chloro-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-1-one

To a mixture of 8-chloro-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-1-one(2.58 g, 9.1 mmol) in chloroform (9.1 mL) and ethyl acetate (9.1 mL) wasadded copper(II) bromide (4.65 g, 19.9 mmol). The reaction was heated to80° C. for 5 hours and then cooled to room temperature. The mixture wasdiluted with dichloromethane and washed twice with a 5:1 solution ofsaturated aqueous ammonium chloride and aqueous ammonium hydroxide(˜28%), and washed once with water. The organic layer was dried withmagnesium sulfate, filtered and concentrated. The crude material waspurified by flash column chromatography (hexanes/DCM) to yield2-bromo-8-chloro-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-1-one (2.45g, 75% yield).

(2S)-1-tert-butyl2-(8-chloro-1-oxo-2,3,4,6-tetrahydro-1H-dibenzo[c,h]chromen-2-yl)pyrrolidine-1,2-dicarboxylate

To a solution of2-bromo-8-chloro-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-1-one (1.05g, 2.9 mmol) and Boc-Pro-OH (1.75 g, 8.1 mmol) in acetonitrile (9.0 mL)was added diisopropylethylamine (1.5 mL, 8.7 mmol). The solution wasstirred under argon at 50° C. for two hours. Extra Boc-Pro-OH (620 mg,2.9 mmol) and diisopropylethylamine (0.5 mL, 2.9 mmol) were added andthe reaction was stirred at 50° C. for 16 hours. The reaction was cooledto room temperature and diluted with ethyl acetate. The organics werewashed with water and brine, dried with magnesium sulfate andconcentrated. The crude material was purified by flash columnchromatography and the product (2S)-1-tert-butyl2-(8-chloro-1-oxo-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-2-yl)pyrrolidine-1,2-dicarboxylate was isolated as a mixture of diastereomers(0.99 g, 69% yield).

tert-butyl(2S)-2-(9-chloro-3,4,5,7-tetrahydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

To a solution of (2S)-1-tert-butyl2-(8-chloro-1-oxo-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-2-yl)pyrrolidine-1,2-dicarboxylate (2.2 g, 4.4 mmol) in toluene (40 mL) wasadded ammonium acetate (7 g, 91 mmol). The reaction mixture wasvigorously refluxed for 3 hours, then cooled to room temperature anddiluted with ethyl acetate. The organics were washed with water andbrine, dried with magnesium sulfate and concentrated. The crude materialwas purified by flash column chromatography to yield tert-butyl(2S)-2-(9-chloro-3,4,5,7-tetrahydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.13 g, 54% yield) as well as recovered (2S)-1-tert-butyl2-(8-chloro-1-oxo-2,3,4,6-tetrahydro-/H-dibenzo[c,h]chromen-2-yl)pyrrolidine-1,2-dicarboxylate (0.8 g, 36%).). LCMS-ESI⁺: calculated forC₂₇H₂₈N₃O₃: 477.98; observed [M+1]⁺: 478.54.

tert-butyl(2S)-2-(9-chloro-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

To a solution of Intermediate tert-butyl(2S)-2-(9-chloro-3,4,5,7-tetrahydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.43 g, 3.0 mmol) in dichloromethane (30 mL) was added manganese(IV)oxide (15 g, 198 mmol). The mixture was stirred for four hours at roomtemperature then filtered through Celite. The MnO₂ was thoroughly rinsedwith dichloromethane and the total filtrate was concentrated to yieldtert-butyl(2S)-2-(9-chloro-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.37 g, 96% yield). This material was used without furtherpurification.

tert-butyl(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate

To a solution of tert-butyl(2S)-2-(9-chloro-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(1.4 g, 2.9 mmol) in dioxane (20 mL) was added bis(pinacolato)diboron(1.5 g, 5.9 mmol), tris(dibenzylideneacetone)dipalladium(0) (110 mg,0.12 mmol), X-Phos (145 mg, 0.30 mmol) and potassium acetate (870 mg,8.9 mmol). The mixture was degassed with a stream of argon for tenminutes. The degassed reaction was heated under argon to 90° C. for 2.5hours then cooled to room temperature and diluted with ethyl acetate.The organics were washed with water and brine, dried with magnesiumsulfate and concentrated. The crude material was purified by flashcolumn chromatography (DCM/EtOAc) to yield tert-butyl(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(1.5 g, 90% yield).

methyl[(2S)-3-methyl-1-oxo-1-{(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}butan-2-yl]carbamate

A solution of tert-butyl(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(0.98 g, 1.7 mmol), concentrated HCl (2 mL) and ethanol (20 mL) washeated to 60° C. for 2 hours. The reaction was concentrated andredissolved in a minimal amount of methanol. An equal volume ofdichloromethane was added and the solution was again concentrated.Dichloromethane was added to the resulting residue and concentrated offtwo more times. The resulting crude material was dissolved indimethylformamide (17 mL). To this solution was added(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (455 mg, 2.6 mmol),HATU (955 mg, 2.5 mmol) and diisopropylethylamine (3 mL, 17 mmol). Thereaction was stirred at room temperature for one hour then diluted withethyl acetate. The organics were washed with water (×2) and brine, driedwith magnesium sulfate and concentrated. The resulting residue waspurified by flash column chromatography to yield Intermediate methyl[(2S)-3-methyl-1-oxo-1-{(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}butan-2-yl]carbamate(780 mg, 72% yield over 2 steps).

tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

A mixture of Pentacyclic Intermediate methyl[(2S)-3-methyl-1-oxo-1-{(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}butan-2-yl]carbamate(780 mg, 1.3 mmol), (S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (450 mg, 1.4mmol), tetrakis(triphenylphosphine)palladium(0) (30 mg, 0.03 mmol),PdCl₂(dppf) (60 mg, 0.08 mmol), 2M aqueous potassium carbonate (1.9 mL,3.9 mmol), dimethoxyethane (10 mL) and dimethylformamide (2 mL) wasdegassed with argon for 15 minutes. The reaction was then heated to 85°C. for 3 hours. Upon completion, the reaction was cooled to roomtemperature, diluted with ethyl acetate and filtered through Celite. Thefiltrate was washed with water and brine, dried (MgSO₄) andconcentrated. The resulting crude material was purified by flash columnchromatography (EtOAc/MeOH) to yield Intermediate tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(390 mg, 43% yield).

methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A mixture of Intermediatetert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(390 mg, 0.53 mmol), concentrated HCl (2 mL) and ethanol (10 mL) washeated to 60° C. for 2 hours. The reaction was concentrated andredissolved in a minimal amount of methanol. An equal volume ofdichloromethane was added and the solution was again concentrated.Dichloromethane was added to the resulting residue and concentrated offtwo more times. One half of the crude material (˜0.27 mmol) wasdissolved in dimethylformamide (2.5 mL). To this solution was added(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (66 mg, 0.38 mmol),HATU (140 mg, 0.37 mmol) and diisopropylethylamine (0.48 mL, 2.7 mmol).The reaction was stirred at room temperature for 2 hours, and thendiluted with acetonitrile (2 mL) and methanol (2 mL). To this solutionwas added ten drops of 5M aqueous NaOH solution and stirring wascontinued for 30 minutes. The reaction was diluted with ethyl acetateand the organic layer was washed with water and brine. The combinedaqueous washings were extracted three times with ethyl acetate, and thecombined organic layers were dried (MgSO₄) and concentrated. The crudematerial was purified by reverse phase HPLC (Gemini, 15 to 45%ACN/H₂O+0.1% TFA) to yield methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(140 mg, 67% yield over 2 steps). LCMS-ESI⁺: calculated for C₄₃H₅₀N₈O₇:790.91; observed [M+1]⁺: 791.71.

Example MA

This compound was made in an analogous manner to methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate,substituting (R)-2-(methoxycarbonylamino)-2-phenylacetic acid for(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid and substituting COMUfor HATU in the final amide coupling step. LCMS-ESI⁺: calculated forC₄₆H₄₈N₈O₇: 824.92; observed [M+1]⁺: 825.72.

Example MB

tert-butyl(2S)-2-[9-(2-{(2R)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate

A mixture of tert-butyl(2S)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(450 mg, 0.79 mmol), methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(325 mg, 0.87 mmol), tetrakis(triphenylphosphine)palladium(0) (30 mg,0.02 mmol), PdCl₂(dppf) (35 mg, 0.05 mmol), 2M aqueous potassiumcarbonate (1.2 mL, 2.4 mmol), dimethoxyethane (6.8 mL) anddimethylformamide (1.2 mL) was degassed with argon for 15 minutes. Thereaction was then heated to 85° C. for 2.5 hours. Upon completion, thereaction was cooled to room temperature, diluted with ethyl acetate andfiltered through Celite. The filtrate was washed with water and brine,dried (MgSO₄) and concentrated. The resulting crude material waspurified by flash column chromatography (EtOAc/MeOH) to yield tert-butyl(2S)-2-[9-(2-{(2R)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate (270 mg, 46%yield).

methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A mixture of tert-butyl(2S)-2-[9-(2-{(2R)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(270 mg, 0.37 mmol), concentrated HCl (1.5 mL) and ethanol (8 mL) washeated to 60° C. for 1 hour. The reaction was concentrated andredissolved in a minimal amount of methanol. An equal volume ofdichloromethane was added and the solution was again concentrated.Dichloromethane was added to the resulting residue and concentrated offtwo more times. The crude material was dissolved in 5:1dichloromethane/dimethylformamide (3.8 mL). To this solution was added(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (96 mg, 0.46 mmol),COMU (190 mg, 0.44 mmol) and diisopropylethylamine (0.20 mL, 1.1 mmol).The reaction was stirred at 0° C. for 30 minutes then warmed to roomtemperature. Upon completion, the reaction was diluted with acetonitrile(2 mL) and methanol (2 mL). To this solution was added ten drops of 5Maqueous NaOH solution and stirring was continued for 30 minutes. Thereaction was diluted with ethyl acetate and the organic layer was washedwith water and brine. The combined aqueous washings were extracted threetimes with ethyl acetate, and the combined organic layers were dried(MgSO₄) and concentrated. The crude material was purified by reversephase HPLC (Gemini, 15 to 45% ACN/H₂O+0.1% TFA) to yield methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(155 mg, 51% yield over 2 steps). LCMS-ESI⁺: calculated for C₄₆H₄₈N₈O₇:824.92; observed [M+1]⁺: 825.67.

Example MC

dimethyl(2S,2′S)-1,1′-((2S,2′S)-2,2′l-pyrrolidin-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-9-yl)-1H-imidazol-2-yl)pyrrolidin-1-yl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate

This compound was made in an analogous manner to methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate,substituting 7-hydroxy-1-tetralone for 5-hydroxy-1-tetralone in thefirst step of the sequence. All reactions in the synthesis gave similarproduct yields as in the synthesis of methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate.LCMS-ESI⁺: calculated for C₄₃H₅₀N₈O₇: 790.91; observed [M+1]⁺: 791.6.

Example MD

This compound was made in an analogous manner to dimethyl(2S,2′S)-1,1′-((2S,2′S)-2,2′l-pyrrolidin-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-9-yl)-1H-imidazol-2-yl)pyrrolidin-1-yl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate,substituting (R)-2-(methoxycarbonylamino)-2-phenylacetic acid for(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid and substituting COMUfor HATU in the final amide coupling step. LCMS-ESI⁺: calculated forC₄₆H₄₈N₈O₇: 824.92; observed [M+1]⁺: 825.67.

Example ME

dimethyl(2S,2′S)-1,1′-((2S,2′S)-2,2′l-pyrrolidin-2-yl)-7H-dihydro-naphtho[c,g]chromeno[8,9-d]imidazol-9-yl)-1H-imidazol-2-yl)pyrrolidin-1-yl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate

This compound was made in an analogous manner to dimethyl(2S,2′S)-1,1′-((2S,2′S)-2,2′1-pyrrolidin-2-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-9-yl)-1H-imidazol-2-yl)pyrrolidin-1-yl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate,omitting the MnO₂ oxidation of tert-butyl2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate.LCMS-ESI⁺: calculated for C₄₃H₅₂N₈O₇: 792.40; observed [M+1]⁺: 793.69.

Example MF

methyl[1-(2-{5-[2-(1-{[(methoxycarbonyl)amino]-3-methyl-1-oxobutan-2-yl}pyrrolidin-2-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl]-1H-imidazol-2-yl}pyrrolidin-1-yl)-phenyl-1-oxoacet-2-yl]carbamate

This compound was made in an analogous manner to dimethyl(2S,2′S)-1,1′-((2S,2′S)-2,2′1-pyrrolidin-2-yl)-7H-dihydro-naphtho[c,g]chromeno[8,9-d]imidazol-9-yl)-1H-imidazol-2-yl)pyrrolidin-1-yl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate,substituting (R)-2-(methoxycarbonylamino)-2-phenylacetic acid for(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid and substituting COMUfor HATU in the final amide coupling step. LCMS-ESI⁺: calculated forC₄₆H₅₀N₈O₇: 826.94; observed [M+1]⁺: 827.71.

Example MG

This compound was made in an analogous manner to methyl{(2S)-1-[(2R)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate,substituting tert-butyl(2S)-2-[9-(2-{(2R)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-3,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylatefor tert-butyl(2S)-2-[9-(2-{(2R)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-3,7-dihydroisochromeno[3′,4′:5,6]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate.LCMS-ESI⁺: calculated for C₄₆H₅₀N₈O₇: 826.94; observed [M+1]⁺: 827.64.

Example MM

(2S,4S)-methyl {4-(methoxymethyl)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]-imidazol-2-yl)pyrrolidine-1-carboxylate(424 mg, 0.69 mmol), ethanol (6 mL) and concentrated HCl (2 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (10 mL). This solution was concentrated and tothis material was added a solution of2-methoxycarbonylamino-3-methylbutyric acid (152 mg, 0.86 mmol) and HATU(303 mg, 0.79 mmol) in DMF (6 mL). To the resulting solution was addeddiisopropylethylamine (360 μL, 2.08 mmol). After stirring for 2 hours atroom temperature, the reaction was diluted with ethyl acetate, washedwith 5% NaHCO₃ solution, water and brine, dried (Na₂SO₄), concentratedand dried under vacuum to give (2S,4S)-methyl{4-(methoxymethyl)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate.

tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

To a solution of(2S,4S)-methyl{4-(methoxymethyl)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.69 mmol), (S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (220 mg, 0.69mmol), tetrakis(triphenylphosphine) palladium(0) (24 mg, 0.02 mmol) anddichloro[1,1′-bis(diphenylphosphino) ferrocene]palladium(II) (31 mg,0.04 mmol) in a mixture of 1,2-dimethoxyethane (6.0 mL) anddimethylformamide (1.0 mL) was added a solution of potassium carbonate(2M in water, 1.04 mL, 2.0 mmol). The resulting mixture was degassed andthen heated to 85° C. under argon for 18 hours. After cooling to roomtemperature, the reaction was diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield (tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(145 mg, 27%).

methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(145 mg, 0.18 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (51 mg, 0.24 mmol) andCOMU (92 mg, 021 mmol) in DMF (3 mL). To the resulting solution wasadded diisopropylethylamine (100 μL, 0.56 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 43% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(68 mg, 39%). MS (ESI) m/z 870 [M+H]⁺. ¹H NMR (400 MHz, dmso) δ 8.71 (s,1H), 8.22 (d, 1H, J=8 Hz), 8.09 (m, 1H), 7.88-7.63 (m, 6H), 7.36-7.29(m, 6H), 5.41 (d, 1H, J=8.4 Hz), 5.30-5.24 (m, 2H), 5.14-5.10 (m, 1H),4.13-3.09 (m, 15H), 2.47-1.80 (m, 8H), 0.80 (dd, 6H, J=6.4 Hz, J=23 Hz).

Example MN

tert-butyl(2S,4S)-2-[9-(2-{(2S4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (438 mg, 0.72 mmol),methyl(S)-1-((2S,4S)-2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(276 mg, 0.72 mmol), tetrakis(triphenylphosphine) palladium(0) (41 mg,0.04 mmol) and dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (52 mg, 0.07 mmol) in a mixture of1,2-dimethoxyethane (8.6 mL) and dimethylformamide (1.5 mL) was added asolution of potassium carbonate (2M in water, 1.07 mL, 2.15 mmol). Theresulting mixture was degassed and then heated to 85° C. under argon for18 hours. After cooling to room temperature, the reaction was dilutedwith ethyl acetate. The organics were washed with water and brine, dried(Na₂SO₄), and concentrated. The crude residue was purified by flashchromatography to yield tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(182 mg, 32%).

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-1-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate (182 mg, 0.18 mmol), ethanol (3 mL) andconcentrated HCl (1 mL) was heated to 60° C. for 1 hour. The reactionwas concentrated and the crude material dissolved in DCM (6 mL). Thissolution was concentrated and to this material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (47 mg, 0.23 mmol) andCOMU (85 mg, 0.2 mmol) in DMF (3 mL). To the resulting solution wasadded diisopropylethylamine (90 μL, 0.52 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 49% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to givemethyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(32 mg, 39%). MS (ESI) m/z 884 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.70 (s, 1H), 8.21 (d, 1H, J=8 Hz), 8.08 (s,1H), 7.90-7.64 (m, 6H), 7.34-7.31 (m, 3H), 7.64 (d, 1H, J=8.4 Hz), 5.47(d, 1H, J=7.6 Hz), 5.28-5.25 (m, 3H), 5.05-5.01 (m, 1H), 4.19-4.04 (m,3H), 3.67-3.15 (m, 15H), 2.51-2.46 (m, 4H), 1.95-1.92 (m, 2H), 1.82-1.76(m, 1H), 1.10 (d, 3H, J=6 Hz), 0.75 (dd, 6H, J=6.8 Hz, J=14 Hz).

Example MO

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(57 mg, 0.08 mmol), 2-methoxycarbonylamino-3-methylbutyric acid (19 mg,0.1 mmol), HATU (303 mg, 0.79 mmol) in DMF (1 mL) was addeddiisopropylethylamine (43 μL, 0.24 mmol). After stirring for 2 hours atroom temperature, the reaction was diluted with ethyl acetate, washedwith 5% NaHCO₃ solution, water and brine, dried (Na₂SO₄), concentratedand purified by preparative reverse phase HPLC (Gemini, 15 to 43%ACN/H₂O+0.1% TFA). The product fractions were lyophilized to give methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate.(13 mg, 19%). MS (ESI) m/z 850 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.66 (s, 1H), 8.28-8.13 (m, 1H), 8.12-7.99 (m,1H), 7.90-7.75 (m, 3H), 7.73-7.65 (m, 1H), 7.63-7.57 (m, 1H), 7.34-7.19(m, 2H), 5.30-5.24 (m, 2H), 5.21-4.95 (m, 2H), 4.33-3.93 (m, 6H),3.23-3.58 (m, 12H), 2.76-2.59 (m, 2H), 2.02-1.73 (m, 6H), 1.12-1.07 (m,3H), 0.86-0.68 (m, 12H).

Example MP

tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(217 mg, 0.35 mmol), methyl(S)-1-((2S,5S)-2-(5-bromo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(170 mg, 0.39 mmol), tetrakis(triphenylphosphine) palladium(0) (21 mg,0.02 mmol) and dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (26 mg, 0.04 mmol) in a mixture of1,2-dimethoxyethane (4.3 mL) and dimethylformamide (0.75 mL) was added asolution of potassium carbonate (2M in water, 0.53 mL, 1.06 mmol). Theresulting mixture was degassed and then heated to 85° C. under argon for18 hours. After cooling to room temperature, the reaction was dilutedwith ethyl acetate. The organics were washed with water and brine, dried(Na₂SO₄), and concentrated. The crude residue was purified by flashchromatography to yield tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate (110 mg, 39%).

methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate (108 mg, 0.14 mmol), ethanol (2 mL) andconcentrated HCl (0.7 mL) was heated to 60° C. for 1 hour. The reactionwas concentrated and the crude material dissolved in DCM (10 mL). Thissolution was concentrated and to this material was added a solution of2-methoxycarbonylamino-3-methylbutyric acid (31 mg, 0.18 mmol) and HATU(60 mg, 0.16 mmol) in DMF (2 mL). To the resulting solution was addeddiisopropylethylamine (70 μL, 0.41 mmol). After stirring for 2 hours atroom temperature, the reaction was diluted with ethyl acetate, washedwith 5% NaHCO₃ solution, water and brine, dried (Na₂SO₄). After stirringfor 2 hours at room temperature, the reaction was diluted with ethylacetate, washed with 5% NaHCO₃ solution, water and brine, dried(Na₂SO₄), concentrated and purified by preparative reverse phase HPLC(Gemini, 15 to 43% ACN/H₂O+0.1% TFA). The product fractions werelyophilized to give methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(52 mg, 45%). MS (ESI) m/z 850 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.69 (s, 1H), 8.18 (d, 1H, J=7.6 Hz), 7.99-7.86(m, 4H), 7.72 (s, 1H), 7.64 (d, 1H, J=8.8 Hz), 7.51 (d, 1H, J=8 Hz),7.23 (d, 1H, J=8.4 Hz), 5.29 (s, 2H), 5.22-5.18 (m, 1H), 5.01-4.70 (m,1H), 4.64-4.61 (m, 1H), 4.21-4.17 (m, 1H), 4.09-4.05 (m, 1H), 3.92-3.88(m, 1H), 3.59-3.08 (m, 14H), 2.67-1.83 (m, 7H), 1.43 (d, 3H, J=6.4 Hz),0.91-0.71 (m, 12H).

Example MQ

(2S,4R)-tert-butyl-4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

A degassed mixture of(2S,4R)-tert-butyl-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (335 mg, 0.64 mmol), bis(pinacolato)diboron(246 mg, 0.96 mmol), potassium acetate (190 mg, 1.9 mmol),tris(dibenzylideneacetone) palladium (24 mg, 0.02 mmol) and2-dicyclohexylphosphino-2′, 4′, 6′-tri-i-propyl-1, 1′-biphenyl (31 mg,0.06 mmol) in 1,4-dioxane (3.3 mL) was heated to 90° C. for 3 hours,cooled to room temperature and diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield(2S,4R)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(379 mg, 96%).

(2S,4R)-tert-butyl-2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4R)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (299 mg, 0.49 mmol), methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(217 mg, 0.58 mmol), tetrakis(triphenylphosphine) palladium(0) (28 mg,0.02 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (35 mg, 0.04mmol) in a mixture of 1,2-dimethoxyethane (4.3 mL) and dimethylformamide(0.75 mL) was added a solution of potassium carbonate (2M in water, 0.73mL, 1.46 mmol). The resulting mixture was degassed and then heated to85° C. under argon for 18 hours. After cooling to room temperature, thereaction was diluted with ethyl acetate. The organics were washed withwater and brine, dried (Na₂SO₄), and concentrated. The crude residue waspurified by flash chromatography to yield (2S,4R)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(170 mg, 45%).

methyl{(1R)-2-[(2S,4R)-2-(9-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-phenylethyl}carbamate

A solution of (2S,4R)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (170 mg, 0.22 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (59 mg, 0.28 mmol) andCOMU (108 mg, 0.25 mmol) in DMF (3 mL). To the resulting solution wasadded diisopropylethylamine (110 μL, 0.66 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 44% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(1R)-2-[(2S,4R)-2-(9-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]-naphtho[1,2-d]imidazol-2-yl}-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-phenylethyl}carbamate (67 mg, 35%). MS (ESI) m/z870 [M+H]⁺. 1H NMR (400 MHz, dmso) δ 8.71 (s, 1H), 8.20 (d, 1H, J=8.4Hz), 8.01 (m, 1H), 7.91-7.64 (m, 6H), 7.38-7.28 (m, 6H), 6.85 (s, 1H),5.51 (d, 1H, J=7.2 Hz), 5.39-5.29 (m, 3H), 5.13-5.09 (m, 1H), 4.11-3.04(m, 15H), 2.77-1.98 (m, 8H), 0.79 (dd, 6H, J=6.8 Hz, J=12.8 Hz).

Example MR

(2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

A degassed mixture of (2S,4S)-tert-butyl2-(9-chloro-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(322 mg, 0.61 mmol), bis(pinacolato) diboron (235 mg, 0.92 mmol),potassium acetate (182 mg, 1.9 mmol),tris(dibenzylideneacetone)palladium (23 mg, 0.02 mmol) and2-dicyclohexylphosphino-2′, 4′, 6′-tri-i-propyl-1, 1′-biphenyl (29 mg,0.06 mmol) in 1,4-dioxane (3.3 mL) was heated to 90° C. for 3 hours,cooled to room temperature and diluted with ethyl acetate. The organicswere washed with water and brine, dried (Na₂SO₄), and concentrated. Thecrude residue was purified by flash chromatography to yield(2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(267 mg, 70%).

(2S,4S)-tert-butyl-2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(267 mg, 0.52 mmol), methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(195 mg, 0.52 mmol), tetrakis (triphenylphosphine) palladium(0) (25 mg,0.02 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (32 mg, 0.04mmol) in a mixture of 1,2-dimethoxyethane (4.3 mL) and dimethylformamide(0.75 mL) was added a solution of potassium carbonate (2M in water, 0.65mL, 1.3 mmol). The resulting mixture was degassed and then heated to 85°C. under argon for 18 hours. After cooling to room temperature, thereaction was diluted with ethyl acetate. The organics were washed withwater and brine, dried (Na₂SO₄), and concentrated. The crude residue waspurified by flash chromatography to yield (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (75 mg, 22%).

methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]-4,5-dihydro-naphtho[1,2-d]imidazol-2-yl}-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of (2S,4S)-tert-butyl 2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl-4,5-dihydro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (75 mg, 0.09 mmol), ethanol (2 mL) andconcentrated HCl (0.6 mL) was heated to 60° C. for 1 hour. The reactionwas concentrated and the crude material dissolved in DCM (6 mL). Thissolution was concentrated and to this material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (26 mg, 0.13 mmol) andCOMU (47 mg, 0.11 mmol) in DMF (2 mL). To the resulting solution wasadded diisopropylethylamine (50 μL, 0.29 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 44% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]-4,5-dihydro-naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(15 mg, 18%).

MS (ESI) m/z 872 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 7.95-7.63 (m, 6H), 7.35-7.25 (m, 7H), 6.97 (s,1H), 5.42 (d, 1H, J=6.8 Hz), 5.18 (s, 2H), 5.09 (s, 2H), 4.28-2.63 (m,19H), 2.47-1.80 (m, 8H), 0.77 (dd, 6H, J=4.8 Hz, J=12.4 Hz).

Example MS

(2S,4S)-tert-butyl-2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)-1,11-dihydroisochromeno14′,3′:6,71 naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (400mg, 0.85 mmol), methyl(S)-1-((S)-2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(360 mg, 0.85 mmol), tetrakis(triphenylphosphine) palladium(0) (38 mg,0.03 mmol) and dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (48 mg, 0.07 mmol) in a mixture of1,2-dimethoxyethane (8.0 mL) and dimethylformamide (1.4 mL) was added asolution of potassium carbonate (2M in water, 0.98 mL, 1.96 mmol). Theresulting mixture was degassed and then heated to 85° C. under argon for18 hours. After cooling to room temperature, the reaction was dilutedwith ethyl acetate. The organics were washed with water and brine, dried(Na₂SO₄), and concentrated. The crude residue was purified by flashchromatography to (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(156 mg, 29%).

methyl{(2S)-2-[[(2S,4S)-2-{9-[2-((2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(156 mg, 0.18 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and to(90 mg, 0.12 mmol) of this material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (34 mg, 0.16 mmol) andCOMU (61 mg, 014 mmol) in DMF (2 mL). To the resulting solution wasadded diisopropylethylamine (60 μL, 0.37 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 49% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(2S)-2-[[(2S,4S)-2-{9-[2-((2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(62 mg, 56%). MS (ESI) m/z 920 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.73 (s, 1H), 8.17 (d, 2H, J=8.4 Hz), 7.94 (d,3H, J=8.8 Hz), 7.84-7.67 (m, 6H), 7.37-7.29 (m, 6H), 5.48 (d, 1H, J=7.6Hz), 5.35-5.20 (m, 5H), 4.14-3.12 (m, 15H), 2.52-1.92 (m, 8H), 0.80 (dd,6H, J=6.8 Hz, J=6.4 Hz).

Example MT

methyl{(2S)-2-[(2S,4S)-2-{9-[2-((2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-y]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of(2S,4S)-tert-butyl-2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(156 mg, 0.18 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (6 mL). This solution was concentrated and to68 mg (0.09 mmol) of this material was added a solution of(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (21 mg, 0.12 mmol)and HATU (41 mg, 0.1 mmol) in DMF (1 mL). To the resulting solution wasadded diisopropylethylamine (50 μL, 0.28 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 44% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(2S)-2-[(2S,4S)-2-{9-[2-((2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-y]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-i-oxobutan-2-yl}carbamate(32 mg, 40%). MS (ESI) m/z 886 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.71 (s, 1H), 8.15 (d, 1H, J=8 Hz), 7.95-7.64(m, 8H), 7.28 (dd, 2H, J=8.8 Hz, J=14.4 Hz), 5.31 (s, 2H), 5.23-5.19 (m,2H), 4.09-3.85 (m, 5H), 3.58-3.28 (m, 14H), 2.47-1.89 (m, 9H), 0.83-0.72(m, 12H).

Example MU

tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of (2S)-methyl{-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-i-oxobutan-2-yl}carbamate (460 mg, 0.74 mmol), (2S,4S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(250 mg, 0.61 mmol), tetrakis (triphenylphosphine) palladium(0) (35 mg,0.03 mmol) and dichloro[1,1′-bis (diphenylphosphino)ferrocene]palladium(II) (45 mg, 0.06 mmol) in a mixture of1,2-dimethoxyethane (9.0 mL) and dimethylformamide (1.5 mL) was added asolution of potassium carbonate (2M in water, 0.92 mL, 1.84 mmol). Theresulting mixture was degassed and then heated to 85° C. under argon for18 hours. After cooling to room temperature, the reaction was dilutedwith ethyl acetate. The organics were washed with water and brine, dried(Na₂SO₄), and concentrated. The crude residue was purified by flashchromatography to tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(123 mg)

methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of tert-butyl (2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-carboxylate(122 mg, 0.16 mmol), ethanol (3 mL) and concentrated HCl (1 mL) washeated to 60° C. for 1 hour. The reaction was concentrated and the crudematerial dissolved in DCM (3 mL). This solution was concentrated and tothis material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (43 mg, 0.2 mmol) andCOMU (77 mg, 018 mmol) in DMF (3 mL). To the resulting solution wasadded diisopropylethylamine (80 μL, 0.37 mmol). After stirring for 2hours at room temperature, the reaction was diluted with ethyl acetate,washed with water and brine, dried (Na₂SO₄), concentrated and purifiedby preparative reverse phase HPLC (Gemini, 15 to 44% ACN/H₂O+0.1% TFA).The product fractions were lyophilized to give methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(60 mg, 44%). MS (ESI) m/z 870 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.71 (s, 1H), 8.22 (d, 1H, J=8 Hz), 8.09 (m,1H), 7.88-7.63 (m, 6H), 7.36-7.29 (m, 6H), 5.41 (d, 1H, J=8.4 Hz),5.30-5.24 (m, 2H), 5.14-5.10 (m, 1H), 4.13-3.09 (m, 15H), 2.47-1.80 (m,8H), 0.80 (dd, 6H, J=6.4 Hz, J=23 Hz).

Example MV

(1R,3S,5R)-tert-butyl-3-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-methoxymethylpyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate

To a solution of(1R,3S,5R)-tert-butyl-3-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (213 mg, 0.37 mmol), methyl(S)-1-((2S,4S)-2-(5-bromo-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(142 mg, 0.31 mmol), tetrakis (triphenylphosphine) palladium(0) (35 mg,0.03 mmol) and dichloro[1,1′-bis (diphenylphosphino)ferrocene]palladium(II) (22 mg, 0.03 mmol) in a mixture of 1,4-dioxane(3.0 mL) and dimethylsulfoxide (3.0 mL) was added a solution ofpotassium carbonate (2M in water, 0.46 mL, 0.9 mmol). The resultingmixture was degassed and then heated to 95° C. under argon for 7 hours.After cooling to room temperature, the reaction was diluted with ethylacetate. The organics were washed with water and brine, dried (Na₂SO₄),and concentrated. The crude residue was purified by flash chromatographyto (1R,3S,5R)-tert-butyl 3-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-methoxymethylpyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (101 mg, 42%).

methyl{(1R)-2-[(1R,3S,5R)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-azabicyclo[3.1.0]hex-3-yl]-2-oxo-1-phenylethyl}carbamate

A solution (1R,3S,5R)-tert-butyl3-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-methoxymethylpyrrolidin-2-yl)-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (101 mg, 0.16 mmol), ethanol (3 mL)and concentrated HCl (1 mL) was heated to 60° C. for 1 hour. Thereaction was concentrated and the crude material dissolved in DCM (3mL). This solution was concentrated and to this material was added asolution of (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (35 mg,0.17 mmol) and COMU (63 mg, 015 mmol) in DMF (3 mL). To the resultingsolution was added diisopropylethylamine (70 μL, 0.38 mmol). Afterstirring for 2 hours at room temperature, the reaction was diluted withethyl acetate, washed with water and brine, dried (Na₂SO₄), concentratedand purified by preparative reverse phase HPLC (Gemini, 15 to 44%ACN/H₂O+0.1% TFA). The product fractions were lyophilized to give methyl{(1R)-2-[(1R,3S,5R)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methoxymethylpyrrolidin-2-yl]-1H-imidazol-5yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1.0]hex-3-yl]-2-oxo-1-phenylethyl}carbamate (71 mg, 63%). MS (ESI)m/z 882 [M+H]⁺.

1H NMR (400 MHz, dmso) δ 8.66 (s, 1H), 8.17 (d, 1H, J=8.8 Hz), 8.04 (s,1H), 7.87-7.59 (m, 6H), 7.39-7.22 (m, 6H), 5.72 (d, 1H, J=7.6 Hz), 5.68(s, 1H), 5.25 (s, 1H), 5.13-5.01 (m, 2H), 4.12-4.00 (m, 2H), 3.81-3.00(m, 13H), 2.60 (m, 1H), 2.43-2.37 (m, 3H), 1.92-1.82 (m, 3H), 0.83-0.58(m, 7H), 0.59 (s, 1H), 0.00 (s, 1H).

Example MW

This compound was synthesized using the same conditions as example 00substituting with the respective(1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hexane-3-carboxylicacid and(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid as appropriate.

MS (ESI) m/z 918 [M+H]⁺.

Example MX

This compound was synthesized using the same conditions as example OOsubstituting with the respective (1R,3S,5R)-2-((2S,3S)-3-methoxy-2(methoxycarbonylamino) butanoyl)-2-azabicyclo[3.1.0]hexane-3-carboxylicacid and (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid asappropriate.

MS (ESI) m/z 898 [M+H]⁺.

Example MY

(2S,4S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate

To a solution of9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (1.32 g,3.63 mmol) in MeCN (40 mL) was added(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid(1.0 g, 4.36 mmol) and DIPEA (0.7 mL, 3.99 mmol). After stirring for 18h, the solution was diluted with EtOAc and washed successively withsaturated aqueous NaHCO₃ and brine. The organics were dried over MgSO₄,filtered and concentrated under reduced pressure. The crude residue waspurified by silica column chromatography (10% to 40% EtOAc/hexanes) toafford (2S,4S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate (1.31 g, 70%).

(2S,4S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

(2S,4S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate (1.31 g, 2.56 mmol) was addedxylenes (25 mL) and ammonium acetate (3.95 g, 51.2 mmol) and thesolution was heated to 136° C. and stirred overnight. The followingmorning, the solution was cooled to rt and was diluted with EtOAc andwashed successively with water, saturated aqueous NaHCO₃ and brine. Theorganics were dried over MgSO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(60% to 100% EtOAc/hexanes) to afford (2S,4S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(711 mg, 56%).

(2S,4S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-(methyl)pyrrolidine-1-carboxylate(935 mg, 1.9 mmol) in CH₂Cl₂ (20 mL) was added MnO₂ (8.25 g, 95 mmol).The reaction mixture was stirred for 3 h, and then filtered over celite.The filter cake was washed with copious CH₂Cl₂ and MeOH, and thefiltrate was concentrated under reduced pressure. The crude residue waspurified by silica column chromatography (0% to 10% MeOH/EtOAc) toafford (2S,4S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(692 mg, 74%).

(2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7] naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate

(2S,4S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(692 mg, 1.41 mmol) in dioxane (15 mL) was added bis(pinacolato)diboron(1.07 g, 4.23 mmol), KOAc (415 mg, 4.23 mmol), X-Phos (52 mg, 0.11mmol), and Pd₂dba₃ (26 mg, 0.03 mmol). The solution was degassed with N2for 10 min, then heated to 100° C. for 16 h. The solution was cooled tort, diluted with EtOAc, washed with saturated aqueous NaHCO₃, brine,dried with MgSO₄, and concentrated. Purified by silica gelchromatography (0% to 30% MeOH/EtOAc) to afford (2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7] naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (821mg, quant).

(2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4(methyl)pyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7] naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (821mg, 1.41 mmol), methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(1.05 g, 2.82 mmol), tetrakis(triphenylphosphine) palladium(0) (162 mg,0.14 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (102 mg,0.14 mmol) in DME (15 mL) was added a solution of potassium carbonate(2M in water, 2.32 mL, 4.65 mmol). The resulting mixture was degassedand then heated to 85° C. for 18 hours. After cooling to roomtemperature, the reaction was diluted with ethyl acetate. The organicswere washed with saturated sodium bicarbonate and brine, dried overMgSO₄ and concentrated. The crude residue was purified by flashchromatography to yield (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4methylpyrrolidine-1-carboxylate(386 mg, 37%).

Methyl{2-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-(methyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of (2S,4S)-tert-butyl2-(9-(2-((S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-4methylpyrrolidine-1-carboxylate(386 mg, 0.52 mmol), CH₂Cl₂ (8 mL), MeOH (2 mL) and HCl (4M in Dioxane,2 mL) and was stirred overnight. The reaction was concentrated and thecrude material dissolved in DMF (8 mL). This solution was concentratedand to this material was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (108 mg, 0.52 mmol) andCOMU (248 mg, 0.52 mmol). To the resulting solution was addeddiisopropylethylamine (0.45 mL, 2.6 mmol). After stirring for 2 hours atroom temperature, the reaction was diluted with 10% MeOH/EtOAc, washedwith saturated NaHCO₃, water and brine, dried (Na₂SO₄), concentrated andpurified by HPLC to give methyl{2-[2-{9-[2-(1-{2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}-4-methylpyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(27 mg, 6%). LCMS-ESI⁺: calculated for C₄₇H₅₀N₈O₇: 838.38; observed[M+1]⁺: 840.12

Example NB

Methyl[(2S)-3-methyl-1-{(2S,4S)-4-methyl-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate

(2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7] naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (950mg, 1.63 mmol) was dissolved in DCM (12 mL), MeOH (3 mL) and HCl (4 M indioxane, 3 mL) was added. The reaction mixture was stirred for 4 h andthen concentrated under reduced pressure. The crude residue was treatedwith (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (285 mg, 1.63mmol), HATU (620 mg, 1.63 mmol) and DMF (15 mL), then DIPEA (1.42 mL,8.15 mmol) was added dropwise. After 1 h, the mixture was diluted withEtOAc and washed successively with saturated aqueous NaHCO₃ and brine.The organics were dried over MgSO₄, filtered and concentrated underreduced pressure. The crude residue was purified by silica columnchromatography (0% to 30% MeOH/EtOAc) to afford methyl[(2S)-3-methyl-1-{(2S,4S)-4-methyl-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(596 mg, 57%).

Tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

Methyl[(2S)-3-methyl-1-{(2S,4S)-4-methyl-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(298 mg, 0.47 mmol), (S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (443 mg, 1.4mmol), Pd(PPh₃)₄ (54 mg, 0.05 mmol), PdCl₂(dppf)₂ (36 mg, 0.05 mmol),and K₂CO₃ (2M in H₂O, 0.78 mL, 1.55 mmol) were combined in DME (5 mL).The mixture was degassed with bubbling N2 for 10 min then heated to 85°C. for 16 h. After cooling, the reaction mixture was diluted with EtOAc,and washed successively with saturated aqueous NaHCO₃ and brine. Theorganics were dried over MgSO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(0% to 30% MeOH/EtOAc) to afford tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(84 mg, 24%).

Methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

Tert-butyl(2S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(84 mg, 0.11 mmol) was dissolved in DCM (2.5 mL), MeOH (0.5 mL) and HCl(4 M in dioxane, 0.5 mL) was added. The reaction mixture was stirred for18 h and then concentrated under reduced pressure. The crude residue wastreated with (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (23 mg,0.11 mmol), COMU (53 mg, 0.11 mmol) and DMF (3 mL), then DIPEA (0.10 mL,0.56 mmol) was added dropwise. After 30 min, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(41 mg, 45%). LCMS-ESI⁺: calculated for C₄₇H₅₀N₈O₇: 838.38; observed[M+1]⁺: 839.39

Example NC

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(164 mg, 0.23 mmol) was dissolved in DCM (2.57 mL), MeOH (0.7 mL) andHCl (4 M in dioxane, 0.7 mL) was added. The reaction mixture was stirredfor 16 h and then concentrated under reduced pressure. The crude residuewas treated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (30mg, 0.17 mmol), HATU (65 mg, 0.17 mmol) and DMF (3 mL), then DIPEA (0.15mL, 0.85 mmol) was added dropwise. After 45 min, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(23 mg, 16%).

LCMS-ESI⁺: calculated for C45H54N8O7: 818.41; observed [M+1]⁺: 820.70.

Example ND

Tert-butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

(2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(293 mg, 0.78 mmol), (2S,4S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate (300 mg,0.52 mmol), Pd(PPh₃)₄ (60 mg, 0.052 mmol), PdCl₂(dppf)₂ (38 mg, 0.052mmol), and K₂CO₃ (2M in H₂O, 0.86 mL, 1.72 mmol) were combined in DME (6mL). The mixture was degassed with bubbling N2 for 10 min then heated to85° C. for 16 h. After cooling, the reaction mixture was diluted withEtOAc, and washed successively with saturated aqueous NaHCO₃ and brine.The organics were dried over MgSO₄, filtered and concentrated underreduced pressure. The crude residue was purified by silica columnchromatography (100% EtOAc) to afford tert-butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(183 mg, 50%).

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(183 mg, 0.26 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4M in dioxane, 1 mL) was added. The reaction mixture was stirred for 2 hand then concentrated under reduced pressure. The crude residue wastreated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (91 mg,0.52 mmol), HATU (198 mg, 0.52 mmol) and DMF (5 mL), then DIPEA (0.45mL, 2.6 mmol) was added dropwise. After 1 h, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl {(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(6 mg, 3%). LCMS-ESI⁺: calculated for C45H54N8O7: 818.41; observed[M+1]⁺: 819.41.

Example NF

Tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

(2S,4S)-tert-butyl4-methyl-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate (558 mg, 0.96mmol), methyl(S)-1-((2S,4S)-2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(501 mg, 1.15 mmol), Pd(PPh₃)₄ (111 mg, 0.096 mmol), PdCl₂(dppf)₂ (70mg, 0.096 mmol), and K₂CO₃ (2M in H₂O, 1.6 mL, 3.17 mmoL) were combinedin DMSO (6 mL) and dioxane (6 mL). The mixture was degassed withbubbling N2 for 10 min then heated to 95° C. for 14 h. After cooling,the reaction mixture was diluted with EtOAc, and washed successivelywith saturated aqueous NaHCO₃ and brine. The organics were dried overMgSO₄, filtered and concentrated under reduced pressure. The cruderesidue was purified by silica column chromatography (0%-30% MeOH/EtOAc)to afford tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(257 mg, 35%).

Methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate (257mg, 0.34 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4 M indioxane, 1 mL) was added. The reaction mixture was stirred for 3 h andthen concentrated under reduced pressure. The crude residue was treatedwith (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (71 mg, 0.34mmol), COMU (161 mg, 0.34 mmol) and DMF (6 mL), then DIPEA (0.3 mL, 1.67mmol) was added dropwise. After 15 h, the mixture was diluted with 10%MeOH/EtOAc and washed successively with saturated aqueous NaHCO₃ andbrine. The organics were dried over MgSO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by HPLC to affordmethyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(152 mg, 53%). LCMS-ESI⁺: calculated for C₄₈H₅₂N₈O₇: 852.40; observed[M+1]⁺: 854.26. ¹H NMR (CD₃OD): 8.677 (s, 1H), 8.232-7.837 (m, 5H),7.695-7.673 (m, 2H), 7.496-7.426 (m, 5H), 5.499 (s, 1H), 5.445-5.401 (m,1H), 5.337 (s, 1H), 5.253-5.208 (q, 1H, J=7.2 Hz), 4.870 (m, 1H), 4.230(d, 1H, J=7.2 Hz), 3.781 (m, 1H), 3.671 (s, 3H), 3.607 (s, 3H), 3.425(m, 3H), 2.750-2.689 (m, 2H), 2.683 (m, 2H), 2.384 (m, 1H), 1.894(quint, 2H, J=12 Hz), 1.249-1.151 (m, 6H), 0.974-0.890 (m, 6H).

Example NG

(2S,5S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)5-methylpyrrolidine-1,2-dicarboxylate

To a solution of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one in MeCN(30 mL) was added(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(1.2 g, 3.23 mmol) and triethyl amine (0.48 mL, 3.55 mmol) and thesolution was heated to 50° C. After stirring for 15 h, the solution wascooled to rt, and diluted with EtOAc and washed successively withsaturated aqueous NaHCO₃ and brine. The organics were dried over MgSO₄,filtered and concentrated under reduced pressure. The crude residue waspurified by silica column chromatography (20% to 50% EtOAc/hexanes) toafford (2S,5S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)5-methylpyrrolidine-1,2-dicarboxylate (1.09 g, 65%).

(2S,5S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate

(2S,5S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)5-methylpyrrolidine-1,2-dicarboxylate (1.29 g, 2.48 mmol) was dissolvedin a solution of DCM (17.5 mL) and MeOH (7 mL), then treated withpyridinium tribromide (873 mg, 2.73 mmol). After stirring at RT for 1 h,the reaction mixture was diluted with DCM and 10% HCl, and extractedwith DCM. The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure and the crude material was carriedon without further purification.

(2S,5S)-2-(2-(9-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate

(2S,5S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (700 mg, 1.17 mmol)was treated with a solution of(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(375 mg, 1.64 mmol) in acetone (6 mL) and Cs₂CO₃ (267 mg, 0.82 mmol).The stirred reaction mixture was heated to 40° C. for 16 h, then cooledto RT and diluted with CH₂Cl₂ and extracted 3×. The organic phase waswashed with brine, then dried over MgSO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (40% to 100% EtOAc/hexanes) to afford(2S,5S)-2-(2-(9-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (464 mg, 53%).

Tert-butyl(2S,5S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

(2S,5S)-2-(2-(9-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (464 mg, 0.62 mmol)and NH₄OAc (8.48 g, 110.0 mmol) were suspended in a solution of 10:1PhMe/2-methoxyethanol (22 mL). The stirred reaction mixture was heatedto 110° C. for 20 h, then cooled to RT and diluted with EtOAc. Theorganic phase was washed with water, saturated aqueous NaHCO₃, andbrine, then dried over MgSO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(0% to 30% MeOH/EtOAc) to afford tert-butyl(2S,5S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(393 mg, 90%).

Tert-butyl(2S,5S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

Tert-butyl(2S,5S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(393 mg, 0.55 mmol) was suspended in DCM (7 mL) and activated MnO₂ (1.45g, 16.7 mmol) was added in a single portion. The reaction mixture washeated to 40° C. After stirring for 2.5 h, the mixture was cooled to rtand the slurry was filtered over celite. The filter cake was washed withcopious CH₂Cl₂ and MeOH and the filtrate was concentrated under reducedpressure. The crude material was taken on to the next step withoutfurther purification to afford tert-butyl(2S,5S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(328 mg, 85%).

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,5S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(164 mg, 0.23 mmol) was dissolved in DCM (7 mL), MeOH (1.5 mL) and HCl(4 M in dioxane, 1.5 mL) was added. The reaction mixture was stirred for16 h and then concentrated under reduced pressure. The crude residue wastreated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (81 mg,0.46 mmol), HATU (175 mg, 0.46 mmol) and DMF (5 mL), then DIPEA (0.4 mL,2.34 mmol) was added dropwise. After 35 min, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(132 mg, 69%). LCMS-ESI⁺: calculated for C45H54N8O7: 818.41; observed[M+1]⁺: 820.19. ¹H NMR (CD₃OD): 8.492 (m, 1H), 8.179-7.538 (m, 7H),5.267-5.201 (m, 3H), 5.125-5.082 (m, 1H), 4.070 (m, 1H), 3.383-3.592 (m,4H), 3.225 (s, 3H), 2.466-2.249 (m, 5H), 1.992-1.892 (m, 3H), 1.568 (d,3H, J=6.4 Hz), 1.490 (d, 3H, J=6.8 Hz), 1.266 (m, 2H), 1.020-0.806 (m,14H).

Example NI

Methyl[(2S,3R)-3-methoxy-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate

Tert-butyl(2S,5S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(164 mg, 0.23 mmol) was dissolved in DCM (7 mL), MeOH (1.5 mL) and HCl(4 M in dioxane, 1.5 mL) was added. The reaction mixture was stirred for16 h and then concentrated under reduced pressure. The crude residue wastreated with (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (90mg, 0.46 mmol), HATU (175 mg, 0.46 mmol) and DMF (6 mL), then DIPEA (0.4mL, 2.34 mmol) was added dropwise. After 30 min, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl[(2S,3R)-3-methoxy-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(97 mg, 50%). LCMS-ESI⁺: calculated for C45H54N8O9: 850.40; observed[M+1]⁺: 851.58. ¹H NMR (CD₃OD): 8.631 (s, 1H), 8.191-7.938 (m, 7H),6.100 (m, 1H), 5.925 (m, 1H), 5.303 (m, 3H), 5.179 (t, 1H, J=6.8 Hz),4.406-4.358 (m, 2H), 3.754-3.598 (m, 8H), 3.376 (s, 3H), 3.263 (s, 3H),2.625-2.256 (m, 6H), 2.038-1.955 (m, 2H), 1.598 (d, 3H, J=6.4 Hz), 1.530(d, 3H, J=6.8 Hz), 1.302-1.099 (m, 6H).

Example NJ

(2S,5S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate

(2S,5S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (800 mg, 1.34 mmol)was treated with a solution of(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (485 mg, 1.87 mmol) in acetone (6 mL) and Cs₂CO₃ (306 mg, 0.94mmol). The stirred reaction mixture was heated to 40° C. for 16 h, thencooled to RT and diluted with CH₂Cl₂ and extracted 3×. The organic phasewas washed with brine, then dried over MgSO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (40% to 100% EtOAc/hexanes) to afford(2S,5S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (680 mg, 65%).

Tert-butyl(2S,5S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

(2S,5S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (680 mg, 0.87 mmol)and NH₄OAc (10.0 g, 130.0 mmol) were suspended in a solution of 10:1PhMe/2-methoxyethanol (22 mL). The stirred reaction mixture was heatedto 110° C. for 24 h, then cooled to RT and diluted with EtOAc. Theorganic phase was washed with water, saturated aqueous NaHCO₃, andbrine, then dried over MgSO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(0% to 30% MeOH/EtOAc) to afford tert-butyl(2S,5S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(461 mg, 72%).

Tert-butyl(2S,5S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

tert-butyl(2S,5S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(461 mg, 0.62 mmol) was suspended in DCM (7 mL) and activated MnO₂ (1.6g, 18.8 mmol) was added in a single portion. The reaction mixture washeated to 40° C. After stirring for 5.5 h, the mixture was cooled to rtand the slurry was filtered over celite. The filter cake was washed withcopious CH₂Cl₂ and MeOH and the filtrate was concentrated under reducedpressure. The crude material was taken on to the next step withoutfurther purification to afford tert-butyl(2S,5S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(414 mg, 90%).

Methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

Tert-butyl(2S,5S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(207 mg, 0.28 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4M in dioxane, 1 mL) was added. The reaction mixture was stirred for 1.5h and then concentrated under reduced pressure. The crude residue wastreated with (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid(106 mg, 0.56 mmol), HATU (214 mg, 0.56 mmol) and DMF (5 mL), then DIPEA(0.49 mL, 2.8 mmol) was added dropwise. After 30 min, the mixture wasdiluted with 10% MeOH/EtOAc and washed successively with saturatedaqueous NaHCO₃ and brine. The organics were dried over MgSO₄, filteredand concentrated under reduced pressure. The crude residue was purifiedby HPLC to afford methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate(132 mg, 69%). LCMS-ESI⁺: calculated for C45H54N8O7: 876.45; observed[M+1]⁺: 879.02

Example NK

(2S,4S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-methylpyrrolidine-1,2-dicarboxylate

To a solution of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (647mg, 1.74 mmol) in MeCN (20 mL) was added((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid(559 mg, 2.44 mmol) and DIPEA (0.36 mL, 2.09 mmol) and the solution washeated to 60° C. After stirring for 3 h, the solution was cooled to rt,and diluted with EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified bysilica column chromatography (20% to 50% EtOAc/hexanes) to afford(2S,4S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-methylpyrrolidine-1,2-dicarboxylate (621 mg, 69%).

(2S,4S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-methylpyrrolidine-1,2-dicarboxylate

(2S,4S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-methylpyrrolidine-1,2-dicarboxylate (621 mg, 1.19 mmol) was dissolvedin a solution of DCM (10 mL) and MeOH (4 mL), then treated withpyridinium tribromide (421 mg, 1.3 mmol). After stirring at RT for 1.5h, the reaction mixture was diluted with DCM and 10% HCl, and extractedwith DCM. The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure and the crude material was carriedon without further purification.

(2S,4S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-methylpyrrolidine-1,2-dicarboxylate

(2S,4S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-methylpyrrolidine-1,2-dicarboxylate (709 mg, 1.18 mmol)was treated with a solution of(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (614 mg, 2.36 mmol) in Me-THF (12 mL) and Cs₂CO₃ (384 mg, 1.18mmol).

The stirred reaction mixture was heated to 50° C. for 16 h, then cooledto RT and diluted with CH₂Cl₂ and extracted 3×. The organic phase waswashed with brine, then dried over MgSO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (40% to 100% EtOAc/hexanes) to afford(2S,4S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-methylpyrrolidine-1,2-dicarboxylate (651 mg, 71%).

Tert-butyl(2S,4S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

(2S,4S)-2-(2-(9-((2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-methylpyrrolidine-1,2-dicarboxylate (651 mg, 0.84 mmol)and NH₄OAc (10.0 g, 129.7 mmol) were suspended in a solution of 10:1PhMe/2-methoxyethanol (22 mL). The stirred reaction mixture was heatedto 110° C. for 20 h, then cooled to RT and diluted with EtOAc. Theorganic phase was washed with water, saturated aqueous NaHCO₃, andbrine, then dried over MgSO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(0% to 30% MeOH/EtOAc) to afford tert-butyl(2S,4S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(382 mg, 62%).

Tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-(methoxymethyl)pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

Tert-butyl(2S,4S)-2-(9-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(382 mg, 0.52 mmol) was suspended in DCM (8 mL) and activated MnO₂ (1.35g, 15.5 mmol) was added in a single portion. The reaction mixture washeated to 35° C. After stirring for 15 h, the mixture was cooled to rtand the slurry was filtered over celite. The filter cake was washed withcopious CH₂Cl₂ and MeOH and the filtrate was concentrated under reducedpressure. The crude material was taken on to the next step withoutfurther purification to afford tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-(methoxymethyl)pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(347 mg, 91%).

Methyl{(2S,3R)-3-methoxy-1-[(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidin-1-yl]-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-(methoxymethyl)pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(174 mg, 0.24 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4M in dioxane, 1 mL) was added. The reaction mixture was stirred for 5 hand then concentrated under reduced pressure. The crude residue wastreated with ((2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid(92 mg, 0.48 mmol), HATU (182 mg, 0.48 mmol) and DMF (5 mL), then DIPEA(0.31 mL, 2.4 mmol) was added dropwise. After 35 min, the mixture wasdiluted with 10% MeOH/EtOAc and washed successively with saturatedaqueous NaHCO₃ and brine. The organics were dried over MgSO₄, filteredand concentrated under reduced pressure. The crude residue was purifiedby HPLC to afford methyl{(2S,3R)-3-methoxy-1-[(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidin-1-yl]-1-oxobutan-2-yl}carbamate(72 mg, 34%). LCMS-ESI⁺: calculated for C46H56N8O10: 880.41; observed[M+1]⁺: 882.39. ¹H NMR (CD₃OD): 8.558 (s, 1H), 8.123-7.572 (m, 7H),5.436-5.391 (dd, 1H, J=7.2, 3.6 Hz), 5.252 (s, 2H), 5.220 (m, 1H),4.493-4.444 (m, 2H), 4.287-4.206 (m, 2H), 3.756-3.256 (m, 21H), 2.834(m, 1H), 2.717-2.621 (m, 2H), 2.500 (m, 1H), 2.150 (m, 1H), 1.882 (m,1H), 1.208 (d, 3H, J=6.4 Hz), 1.159-1.099 (m, 6H).

Example NL

(2S,5S)-2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate

To a solution of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (750mg, 2.02 mmol) in MeCN (20 mL) was added(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid (600 mg, 2.09 mmol) and DIPEA (0.35 mL, 2.02 mmol) and the solutionwas heated to 60° C. After stirring for 4 h, the solution was cooled tort, and diluted with EtOAc and washed successively with saturatedaqueous NaHCO₃ and brine. The organics were dried over MgSO₄, filteredand concentrated under reduced pressure. The crude residue was purifiedby silica column chromatography (40% to 80% EtOAc/hexanes) to afford(2S,5S)-2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate(1.16 g, quant.).

(2S,5S)-2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate

(2S,5S)-2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate(400 mg, 0.61 mmol) was dissolved in a solution of DCM (15 mL) and MeOH(6 mL), then treated with pyridinium tribromide (409 mg, 1.28 mmol). At2 h, an additional portion of pyridinium tribromide (40 mg) was added.After stirring at RT for another 20 min, the reaction mixture wasdiluted with DCM and 10% HCl, and extracted with DCM. The organic phasewas dried over MgSO₄, filtered and concentrated under reduced pressureand the crude material was carried on without further purification.

(2S,4S)-1-tert-butyl2-(3-(2-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate

(2S,5S)-2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate)was treated with a solution of(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid(280 mg, 1.22 mmol) in Me-THF (6 mL) and Cs₂CO₃ (199 mg, 0.61 mmol). Thestirred reaction mixture was heated to 50° C. for 2.5 h, then cooled toRT and diluted with CH₂Cl₂ and extracted 3×. The organic phase waswashed with brine, then dried over MgSO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (50% to 100% EtOAc/hexanes) to afford(2S,4S)-1-tert-butyl 2-(3-(2-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate (441 mg, 90%).

Tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

(2S,4S)-1-tert-butyl2-(3-(2-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-methylpyrrolidine-1,2-dicarboxylate (441 mg, 0.55 mmol) and NH₄OAc (5g, 65.0 mmol) were suspended in a solution of 10:1 PhMe/2-methoxyethanol(11 mL). The stirred reaction mixture was heated to 110° C. for 7 h,then cooled to RT and diluted with EtOAc. The organic phase was washedwith water, saturated aqueous NaHCO₃, and brine, then dried over MgSO₄,filtered and concentrated under reduced pressure. The crude residue waspurified by silica column chromatography (0% to 30% MeOH/EtOAc) toafford tert-butyl (2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(266 mg, 63%).

Tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

Tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(266 mg, 0.35 mmol) was suspended in DCM (7 mL) and activated MnO₂ (908mg, 10.45 mmol) was added in a single portion. The reaction mixture wasstirred overnight. After stirring for 15 h, additional activated MnO₂(500 mg, 5.75 mmol) was added in a single portion.

After stirring 2 h at 35° C., the mixture was cooled to rt and theslurry was filtered over celite. The filter cake was washed with copiousCH₂Cl₂ and MeOH and the filtrate was concentrated under reducedpressure. The crude material was taken on to the next step withoutfurther purification to afford tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(266 mg, quant).

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(266 mg, 0.23 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4M in dioxane, 1 mL) was added. The reaction mixture was stirred for 1.5h and then concentrated under reduced pressure. The crude residue wastreated with (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (44mg, 0.23 mmol), HATU (87 mg, 0.23 mmol) and DMF (5 mL), then DIPEA (0.3mL, 1.75 mmol) was added dropwise. After 30 min, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(59 mg, 31%). LCMS-ESI⁺: calculated for C45H54N8O8: 834.41; observed[M+1]⁺: 836.89. ¹H NMR (CD₃OD): 8.186 (s, 1H), 7.800-7.291 (m, 7H),5.258-5.213 (dd, 1H, J=7.2, 3.6 Hz), 5.027-4.918 (m, 4H), 4.620 (t, 1H,J=6.8 Hz), 4.246 (m, 1H), 4.116 (m, 1H), 3.972 (d, 1H, J=8.8 Hz),3.701-3.675 (m, 1H), 3.503 (s, 3H), 3.479 (s, 3H), 3.177 (s, 3H),2.554-2.191 (m, 3H), 1.906-1.821 (m, 6H), 1.392 (d, 2H, J=6.4 Hz),1.113-0.728 (m, 12H).

Example NM

Tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

Methyl[(2S)-3-methyl-1-{(2S,4S)-4-methyl-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(312 mg, 0.49 mmol), methyl(S)-1-((2S,4S)-2-(5-iodo-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(219 mg, 0.54 mmol), Pd(PPh₃)₄ (58 mg, 0.05 mmol), PdCl₂(dppf)₂ (36 mg,0.05 mmol), and K₂CO₃ (2M in H₂O, 0.8 mL, 1.6 mmoL) were combined inDMSO (5 mL) and dioxane (5 mL). The mixture was degassed with bubblingN2 for 10 min then heated to 95° C. for 5 h. After cooling, the reactionmixture was diluted with EtOAc, and washed successively with saturatedaqueous NaHCO₃ and brine. The organics were dried over MgSO₄, filteredand concentrated under reduced pressure. The crude residue was purifiedby silica column chromatography (0%-30% MeOH/EtOAc) to afford tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(166 mg, 43%).

Methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

Tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(166 mg, 0.21 mmol) was dissolved in DCM (4 mL), MeOH (1 mL) and HCl (4M in dioxane, 1 mL) was added. The reaction mixture was stirred for 2 hand then concentrated under reduced pressure. The crude residue wastreated with (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (44 mg,0.21 mmol), COMU (100 mg, 0.21 mmol) and DMF (5 mL), then DIPEA (0.18mL, 1.05 mmol) was added dropwise. After 1 h, the mixture was dilutedwith 10% MeOH/EtOAc and washed successively with saturated aqueousNaHCO₃ and brine. The organics were dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byHPLC to afford methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(71 mg, 38%). LCMS-ESI⁺: calculated for C₄₉H₅₄N₈O₈: 882.41; observed[M+1]⁺: 884.34. ¹H NMR (CD₃OD): 8.462 (s, 1H), 8.029-7.471 (m, 7H),7.394-7.343 (m, 5H), 5.410 (d, 2H, J=6.8 Hz), 5.300 (m, 1H), 5.233 (m,2H), 4.341 (m, 1H), 4.236 (d, 1H, J=7.2 Hz), 3.603 (s, 3H), 3.551 (s,3H), 3.522-3.241 (m, 8H), 2.650 (m, 1H), 2.550 (m, 2H), 1.977-1.926 (m,4H), 1.221 (d, 3H, J=3.2 Hz), 0.897-0.779 (dd, 6H, J=19.2, 6.8 Hz).

Example NO

(S)-ethyl 2-(tert-butoxycarbonylamino)-5-oxohexanoate

A solution of ethyl N-Boc (S)-pyroglutamate (20.0 g, 77.7 mmol) was inanhydrous THF (150 mL) in a two neck round bottom under argon was cooledto −40° C. Methyl-magnesium bromide solution (3.0 M in Ether, 28.5 mL,85.5 mmol) was added to the reaction mixture dropwise over 30 minutes.The reaction was stirred for 4 hrs at −40° C. then for 1 hr at 0° C. Thereaction was partitioned between ethyl acetate and saturated ammoniumchloride solution and acidified with 1 N HCl. The aqueous layer wasextracted two more times with ethylacetate. The organic layers werecombined and dried with sodium sulfate. The crude material was purifiedby column chromatography (20%-40% EtOAc/hexanes) to yield (S)-ethyl2-(tert-butoxycarbonylamino)-5-oxohexanoate as a viscous oil and wasused directly in the following step.

(S)-ethyl 5-methyl-3,4-dihydro-2H-pyrrole-2-carboxylate

(S)-ethyl 2-(tert-butoxycarbonylamino)-5-oxohexanoate in a 1 L flask wastreated with a trifluoro acetic acid/dichloromethane solution (1:1mixture, 100 mL). Effervescence was observed and the mixture was allowedto stir for 4 hours at room temperature. After which time the volatileswere removed in vacuo to yield (S)-ethyl5-methyl-3,4-dihydro-2H-pyrrole-2-carboxylate as an oil, and useddirectly in the following step.

(2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate

The crude imine in a 1 L flask was dissolved with ethanol (400 mL) wasevacuated and charged with argon three times (3×). Palladium on carbon(apprx. 750 mg, 10% w/w, dry) was added and the reaction was evacuatedof gas and charged with hydrogen gas (3×).

The reaction was allowed to stir under atmospheric hydrogen for 16hours. The mixture was filtered through a plug of celite and thefiltrate was concentrated in vacuo. Diethyl ether was added to the oiland a precipitate formed. The mixture was filtered to yield(2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate, as a white solid (10.6g, 67.4 mmol, 86.7% over three steps). ¹H NMR (400 MHz, cdcl₃) δ 4.48(dd, 1H), 4.27 (q, 2H), 3.92-3.80 (m, 1H), 2.52-2.36 (m, 1H), 2.32-2.13(m, 2H), 1.75-1.60 (m, 1H), 1.51 (d, 3H), 1.30 (t, 3H).

(2S,5S)-1-tert-butyl 2-ethyl 5-methylpyrrolidine-1,2-dicarboxylate

To a solution of (2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate (7.0 g,44.5 mmol) in dichloromethane (250 mL), ditertbutylanhydride (10.7 g,49.0 mmol), diisopropylethylamine (17.1 mL, 98.0 mmol) dropwise over 10minutes, and dimethyl amino pyridine (0.27 g, 2.23 mmol) were addedsuccessively. Effervescence was observed and the mixture was allowed tostir for 16 hours at room temperature. The reaction was washed with HCl(250 mL, of 1N). The organic layer was then dried with sodium sulfate.The crude material was purified by column chromatography (5%-25%EtOAc/hexanes) to yield (2S,5S)-1-tert-butyl 2-ethyl5-methylpyrrolidine-1,2-dicarboxylate as an oil (6.46 g, 25.1 mmol,56%). LCMS-ESI⁺: calc'd for C₁₃H₂₃NO₄: 257.16 (M⁺); Found: 258.70(M+H⁺).

(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid

To a solution of (2S,5S)-1-tert-butyl 2-ethyl5-methylpyrrolidine-1,2-dicarboxylate (6.46 g, 25.1 mmol) in ethanol (20mL) was added lithium hydroxide mono hydrate (2.11 g, 50.2 mmol) anddeionized water (12 mL). The mixture was allowed to stir for 16 hoursthen partitioned between ethylacetate and a 1:1 mixture of saturatedbrine and 1N HCl. The aqueous layer was extracted an additional timewith ethyl acetate. The organic layers were combined, dried with sodiumsulfate and the solvent was removed in vacuo to yield(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid asa white solid (quant.) and was used directly in the following step.

(2S,5S)-tert-butyl 2-(hydroxymethyl)-5-methylpyrrolidine-1-carboxylate

To a solution of(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(5.91 g, 25.8 mmol) in tetrahydrofuran at 0° C., was added borane indimethylsulfide (1.0 M, 3.4 mL, 34 mmol) dropwise. The reaction wasstirred for 4 hours at 0° C. then 18 hours at room temperature. Themixture was then cooled to 0° C. and methanol (70 mL) was addeddropwise. The reaction was warmed to room temperature and the solventswere removed in vacuo. The residue was taken up in dichloromethane (200mL) and extracted with saturated sodium bicarbonate. The organic layerwas dried with sodium sulfate and the solvent was removed in vacuo toyield (2S,5S)-tert-butyl2-(hydroxymethyl)-5-methylpyrrolidine-1-carboxylate as a clear oil (5.15g, 23.9 mmol, 93%) and was used directly in the following step.

(2S,5S)-tert-butyl 2-formyl-5-methylpyrrolidine-1-carboxylate

To a solution of (2S,5S)-tert-butyl2-(hydroxymethyl)-5-methylpyrrolidine-1-carboxylate (5.15 g, 23.9 mmol)in dichloromethane, was added TEMPO (0.075 g, 0.48 mmol), sodium bromide(0.246 g, 2.39 mmol) and sodium bicarbonate (0.442 g, 5.26 mmol). Sodiumhypochlorite (2.67 g, 35.9 mmol) of a 6% solution was added and thebiphasic mixture was vigorously stirred for 2 hours at room temperature.The reaction mixture was extracted two times with dichloromethane (2×100mL). The organic layers were combined and washed with saturated sodiumthiosulfate solution, dried with sodium sulfate and the solvent wasremoved in vacuo to yield (2S,5S)-tert-butyl2-formyl-5-methylpyrrolidine-1-carboxylate (3.9 g, 18.29 mmol, 77%) as aslight colored oil and was used directly in the following step.

(2S,5S)-tert-butyl2-(1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

To a solution of (2S,5 S)-tert-butyl2-formyl-5-methylpyrrolidine-1-carboxylate (3.9g, 18.30 mmol) in MeOH(15 mL) and ammonium hydroxide (15 mL, 99.9%), glyoxal (11.7 mL, 40% w/vin water, 102.40 mmol) was added dropwise. The biphasic mixture turnedorange and turbid. The reaction was stirred vigorously overnight at roomtemperature. The solvent was removed in vacuo. The crude mixture wasredissolved in ethyl acetate and washed with water. The aqueous layerwas washed an additional time with ethyl acetate. The organic layerswere combined and washed with brine, dried with sodium sulfate and thesolvent was removed in vacuo. The crude material was purified by columnchromatography 85% to 100% ethyl acetate in hexanes to yield(2S,5S)-tert-butyl2-(1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate as an off whitesolid (3.47 g, 13.8 mmol, 75%). LCMS-ESI⁺: calc'd for C₁₃H₂₁N₃O₂: 251.16(M+); Found: 252.20 (M+H⁺).

(2S,5S)-tert-butyl2-(4,5-diiodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

A 500 mL round bottom flask was charged with (2S,5S)-tert-butyl2-(1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate (3.47 g, 13.8mmol), iodine (7.7 g, 30.4 mmol) and sodium carbonate (4.54 g, 42.8mmol). Dioxane (70 mL) and water (45 mL) was added to mixture and thereaction was stirred vigorously overnight in the dark. The reaction wasthen partitioned between ethyl acetate and a 10% aqueous solution ofsodium thiosulfate and extracted. The aqueous layer was extracted anadditional time with ethyl acetate. The organic layers were combined,dried with sodium sulfate and the solvent was removed in vacuo. Thecrude material was filtered through a plug of silica with 25% ethylacetate in hexanes to yield (2S,5S)-tert-butyl2-(4,5-diiodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate as awhite solid (4.28 g, 8.50 mmol, 62%). LCMS-ESI⁺: calc'd forC₁₃H₁₉I₂N₃O₂: 502.96 (M+); Found: 503.94 (M+H⁺).

(2S,5S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

To a solution of (2S,5S)-tert-butyl2-(4,5-diiodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate (4.28g, 8.50 mmol) in ethanol (75 mL) and water (75 mL), sodium thiosulfate(10.72 g, 85.1 mmol) was added and the reaction mixture was stirredvigorously for 1 hour at 100° C., 16 hours at 90° C., and 5 hours at100° C. The reaction mixture was partitioned between ethyl acetate andwater. The aqueous layer was washed additionally with ethyl acetate andthe organic layers were combined. The organic layer was dried withsodium sulfate, concentrated and the crude material was purified bycolumn chromatography to yield (2S,5S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate as a whitesolid (2.34 g, 6.20 mmol, 73%). ¹H NMR (400 MHz, cdcl₃) δ 7.04 (s, 1H),4.89 (dd, 1H), 3.92 (m, 1H), 2.91 (s, 1H), 2.18-2.06 (m, 2H), 1.78 (m,1H), 1.52 (m, 1H), 1.48 (s, 9H), 1.13 (d, 3H).

(2S)-1-[(2S,5S)-2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-one

A round bottom flask was charged with (2S,5S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate (1.5 g,3.98 mmol) and treated with an excess of hydrochloric acid (100 mL of4.0M in dioxane). The mixture was stirred vigorously for 3 hours inwhich time a precipitate formed and the solvent was removed in vacuo. Toa mixture of the crude intermediate,(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.836 g, 4.77 mmol),HATU (1.81 g, 4.77 mmol) in dichloromethane (25 mL),diisopropylethylamine (3.46 mL, 19.9 mmol) was then added dropwise andwas stirred over night under nitrogen. The reaction mixture waspartitioned ethyl acetate and saturated sodium bicarbonate. The organiclayer was dried with sodium sulfate, the solvent removed in vacuo. Thecrude product was purified by column chromatography to yield(2S)-1-[(2S,5S)-2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-oneas a white solid (1.63 g, 3.75 mmol, 94%). LCMS-ESI⁺: calc'd forC15H231N403: 434.08 (M⁺); Found: 435.51 (M+H⁺).

Example NP

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

The synthesis of this compound was prepared according to the procedureof example LR-1 with the following modification. During the Suzukicoupling,(2S)-1-[(2S,5S)-2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-onewas used in lieu of(2S)-1-[(2S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-one.The crude material was purified by preparative HPLC to provide methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11 dihydroisochromeno [4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate as a white solid (17 mg, 0.019 mmol, 17%). ¹H NMR (400 MHz,cd₃od) δ 8.63 (s, 1H), 8.19 (d, 1H), 8.04 (m, 1H), 7.87 (m, 2H), 7.66(m, 2H), 7.52-7.39 (m, 6H), 5.50 (m, 2H), 5.32 (s, 2H), 5.16 (m, 1H),4.12 (m, 1H), 3.80 (m, 4H), 3.66 (s, 6H), 3.43 (m, 4H), 3.23 (s, 3H),2.72-1.99 (m, 9H), 1.56 (d, 3H), 1.29 (m, 1H), 0.99 (d, 3H), 0.88 (d,3H).

Example NQ

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

The synthesis of this compound was prepared according to the procedureof example LQ with the following modification. During the Suzukicoupling,(2S)-1-[(2S,5S)-2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-onewas used in lieu of(2S)-1-[(2S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-[(1-methoxyethenyl)amino]-3-methylbutan-1-one.The crude material was purified by preparative HPLC to provide methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateas a white solid (110 mg, 0.131 mmol, 57%). ¹H NMR (400 MHz, cd₃od) δ8.65 (s, 1H), 8.21 (d, 1H), 8.04 (m, 2H), 7.91 (s, 1H), 7.81 (m, 1H),7.67 (m, 2H), 7.46 (m, 6H), 5.59 (s, 1H), 5.50 (dd, 1H), 5.33 (s, 2H),5.22-5.09 (m, 1H), 4.14 (m, 2H), 3.74 (s, 1H), 3.65 (m, 6H), 3.52-3.37(m, 2H), 2.60-1.89 (m, 11H), 1.56 (d, 3H), 1.29 (d, 1H), 0.99 (d, 3H),0.88 (d, 3H).

Example NR Methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S)-1-[(2R)-2-amino-2-phenylacetyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

The synthesis of this compound was prepared according to Example NQ withthe following modifications. During the amide coupling,(R)-2-(tert-butoxycarbonylamino)-2-phenylacetic acid was used in lieu of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid. This was then treatedwith an excess of hydrochloric acid (15 mL, 4.0 M in Dioxane) for 2hours. The crude product was purified by HPLC to provide methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S)-1-[(2R)-2-amino-2-phenylacetyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamateas a white solid (153 mg, 0.196 mmol, 74%). ¹H NMR (400 MHz, cd₃od) δ8.63 (s, 1H), 8.20 (d, 1H), 7.99 (m, 1H), 7.93 (m, 2H), 7.80 (m, 2H),7.72-7.64 (m, 2H), 7.63-7.52 (m, 5H), 5.52 (dd, 1H), 5.44 (m, 1H), 5.33(s, 2H), 5.21-5.10 (m, 1H), 4.80 (m, 2H), 4.14 (m, 1H), 4.02 (m, 1H),3.75 (s, 1H), 3.67 (s, 3H), 3.12 (dd, 1H), 2.72-2.13 (m, 7H), 2.00 (m,3H), 1.56 (d, 3H), 1.30 (d, 1H), 0.98 (d, 3H), 0.88 (d, 3H).

Example OE

(2S,5S)-Ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate

(2S,5S)-Ethyl 5-methylpyrrolidine-2-carboxylate-TFA (10.0 g, 39.3 mmol),(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (6.88 g, 39.3 mmol)and HATU (14.9 g, 39.3 mmol) were combined in DMF (100 mL) and DIPEA(15.0 mL, 86.5 mmol) was added. After stirring for 1 h at RT, thereaction mixture was diluted with EtOAc. The organic phase was washedsuccessively with 10% HCl, saturated aqueous NaHCO₃ and brine, thendried over MgSO₄, filtered and concentrated under reduced pressure toafford (2S,5S)-ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate.The crude material was carried on without further purification.

(2S,5S)-1-((S)-2-(Methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicAcid

(2S,5S)-Ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate(39.3 mmol, assuming complete conversion from the previoustransformation) was suspended in MeOH (200 mL) and aqueous LiOH (1.0 M,100 mL, 100 mmol) was added. The reaction mixture was stirred o/n, thenconcentrated under reduced pressure to remove most of the MeOH. Theaqueous solution was washed 2× with DCM before being acidified to pH-1-2with 10% HCl. The acidic aqueous phase was then extracted 5× with EtOAc.The combined EtOAc extracts were dried over MgSO₄ filtered andconcentrated under reduced pressure to afford(2S,5S)-1-((S)-2-(Methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid (6.89 g, 56% over 2 steps).

Example OF

3-Vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 3-neck oven-dried 500 mL round-bottom flask was cooled under Ar, thencharged with 3-Chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(12.0 g, 42.1 mmol), potassium vinyltrifluoroborate (8.47 g, 6.32 mmol),Pd(OAc)₂ (473 mg, 2.11 mmol), SPhos (1.74 g, 4.25 mmol), K₂C03 (17.5 g,126 mmol) and anhydrous propanol (120 mL). The reaction mixture wassparged with Ar for 16 min, then heated to reflux for 5.5 h. Uponcompletion, the reaction mixture was cooled to RT and concentrated underreduced pressure. The crude residue was suspended in DCM, then washedwith H₂O and brine. The organic solution was dried over MgSO₄, filteredand concentrated under reduced pressure. The resulting residue wasfurther purified via silica plug, eluting with DCM to afford3-vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (10.2 g, 87%).

3-(2-Bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

3-Vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (9.98 g, 36.1mmol) was dissolved in a stirred solution of THF (70 mL), DMSO (70 mL)and H₂O (35 mL). NBS (6.75 g, 37.9 mmol) was added in a single portionand the reaction mixture was stirred at RT for 33 min. Upon completion,the reaction medium was diluted with EtOAc and washed twice with H₂O andonce with brine. The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure. The resulting crude bromohydrin wassuspended in DCM (200 mL) and treated with activated MnO₂ (62.7 g, 722mmol). After stirring for 15 h at RT, the reaction mixture was filteredover celite and the filter cake was rinsed several times with DCM. Thecombined filtrate (˜400 mL) was treated with MeOH (˜100 mL) and themixture was gradually concentrated under reduced pressure, causing solidmaterial to precipitate from solution. When the liquid volume reached˜200 mL, the solid was filtered off and rinsed with MeOH. Theconcentration/precipitation/filtration/rinsing sequence was performed 2×more, resulting in the collection of 3 crops of powdered3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (7.49g, 56% over 2 steps).

(2S,4S)-1-tert-Butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate

3-(2-Bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (7.47g, 20.1 mmol) and(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (5.22 g, 20.1 mmol) were suspended in 2-Me-THF (75 mL) and treatedwith Cs₂CO₃ (3.27 g, 10.1 mmol). After stirring 4 h at RT, the reactionmixture was diluted with DCM. The organic layer was washed with H₂O. Theaqueous layer was then back extracted 2× with DCM. The combined organicswere dried over MgSO₄, filtered and concentrated under reduced pressure.The crude residue was purified by silica column chromatography (10% to50% EtOAc/DCM) to afford (2S,4S)-1-tert-butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (7.73 g, 70%).

(2S,4S)-2-(2-(9-Bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen3-yl)-2-oxoethyl)1-tert-butyl 4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate

(2S,4S)-1-tert-Butyl2-(2-oxo-2-(8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)ethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (7.66 g, 13.9 mmol) wasdissolved in a solution of DCM (100 mL) and MeOH (40 mL), then treatedwith pyridinium tribromide (4.90 g, 15.3 mmol). After stirring at RT for1.75 h, the reaction mixture was diluted with DCM and washedsuccessively with 10% HCl, saturated aqueous NaHCO₃ and brine. Theorganic phase was dried over MgSO₄, filtered and concentrated underreduced pressure and the crude material was carried on without furtherpurification.

(2S,4S)-1-tert-Butyl2-(2-(9-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,91,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate

(2S,4S)-2-(2-(9-Bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (8.76 g,13.94 mmol) was treated with a solution of(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid (6.85 g, 23.92 mmol) in 2-Me-THF (70 mL) and Cs₂CO₃ (3.63 g, 11.15mmol). The stirred reaction mixture was heated to 50° C. for 20 h, thencooled to RT and diluted with EtOAc. The organic phase was washed withH₂O and brine, then dried over MgSO₄, filtered and concentrated underreduced pressure. The crude residue was purified by silica columnchromatography (0% to 30% MeOH/EtOAc) to afford (2S,4S)-1-tert-butyl2-(2-(9-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (10.47 g, 90%).

tert-Butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

(2S,4S)-1-tert-Butyl2-(2-(9-((2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carbonyloxy)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (10.47 g, 12.56 mmol) andNH₄OAc (50.9 g, 660 mmol) were suspended in a solution of 10:1PhMe/2-methoxyethanol (132 mL). The stirred reaction mixture was heatedto 110° C. for 4.5 h, then cooled to RT and diluted with EtOAc. Theorganic phase was washed 3× with saturated aqueous NaHCO₃, then driedover MgSO₄, filtered and concentrated under reduced pressure. The cruderesidue was purified by silica column chromatography (0% to 30%MeOH/EtOAc) to afford tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(8.33 g, 84%).

tert-Butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

tert-Butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(8.33 g, 1.049 mmol) was suspended in DCM and activated MnO₂ (55.0 g,630 mmol) was added in a single portion. After 13 h, MeOH (200 mL) wasadded and the slurry was filtered over celite. The filter cake waswashed with MeOH (600 mL) and the filtrate was concentrated underreduced pressure. The crude material was purified by silica columnchromatography (0% to 45% MeOH/EtOAc) to afford tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(4.85 g, 58%).

Methyl{(2S,3S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

tert-Butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(179 mg, 0.226 mmol) was dissolved in DCM (4 mL) and HCl (4.0 M indioxane, 1 mL) was added. The reaction mixture was stirred for 1 h at RTthen concentrated under reduced pressure. The resulting residue wastreated with (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid (51mg, 0.27 mmol), HATU (95 mg, 0.25 mmol), DMF (2 mL) and DIPEA (0.39 mL,2.3 mmol). After stirring for 6 min, the reaction was quenched with H₂O,filtered and purified by reverse phase HPLC to afford methyl{(2S,3S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate (116 mg, 59%). MS (ESI) m/z 864 [M+H]⁺. ¹H NMR (400 MHz,cd₃od) δ 8.57 (d, J=14.7 Hz, 1H), 8.45 (s, 1H), 8.20 (d, J=14.4 Hz, 1H),8.15-7.98 (m, 2H), 7.91 (dd, J=21.8, 14.1 Hz, 2H), 7.85-7.69 (m, 2H),7.69-7.48 (m, 2H), 5.42-5.12 (m, 5H), 4.34 (dd, J=22.3, 13.7 Hz, 1H),4.30-4.10 (m, 2H), 3.87-3.73 (m, 1H), 3.73-3.63 (m, 7H), 3.62-3.48 (m,2H), 3.48-3.38 (m, 4H), 3.35 (s, 3H), 2.95-2.70 (m, 1H), 2.70-2.55 (m,2H), 2.55-2.20 (m, 2H), 2.20-1.91 (m, 3H), 1.77 (d, J=42.0 Hz, 1H), 1.65(d, J=6.6 Hz, 3H), 1.43 (t, J=24.6 Hz, 1H), 1.28 (d, J=6.2 Hz, 1H),1.23-1.01 (m, 3H), 0.98 (d, J=6.6 Hz, 3H), 0.90 (dd, J=13.1, 5.9 Hz,10H).

Example OG

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate was prepared from tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylateby the same method employed in the synthesis of methyl{(2S,3S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate, replacing (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoicacid with (2S,3R)-2-(methoxycarbonylamino)-3-methylpentanoic acid. MS(ESI) m/z 864 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.62-8.41 (m, 1H), 8.22(s, 1H), 8.07 (dt, J=20.1, 10.0 Hz, 1H), 7.89 (dt, J=35.6, 15.6 Hz, 2H),7.77 (dd, J=20.3, 7.0 Hz, 2H), 7.68-7.48 (m, 2H), 5.95 (d, J=5.0 Hz,1H), 5.42-5.13 (m, 4H), 4.47 (t, J=5.5 Hz, 1H), 4.40-4.09 (m, 2H),3.80-3.73 (m, 1H), 3.73-3.62 (m, 6H), 3.57 (dt, J=16.1, 9.7 Hz, 2H),3.40 (s, 3H), 3.34 (d, J=7.5 Hz, 1H), 2.81 (dd, J=18.4, 12.5 Hz, 1H),2.63 (td, J=13.3, 6.8 Hz, 2H), 2.55-2.18 (m, 2H), 2.16-1.77 (m, 4H),1.65 (d, J=6.6 Hz, 3H), 1.50-1.31 (m, 1H), 1.26 (dd, J=15.6, 6.7 Hz,2H), 1.17-1.03 (m, 2H), 0.98 (dd, J=6.7, 4.5 Hz, 5H), 0.89 (dd, J=15.5,7.8 Hz, 3H), 0.86-0.74 (m, 3H).

Example OH

Methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

tert-Butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(102 mg, 0.128 mmol) was dissolved in DCM (4 mL) and HCl (4.0 M indioxane, 2.0 mL, 8.0 mmol) was added. After stirring at RT for 30 min,the solution was concentrated under reduced pressure. The residue wastreated with (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (29 mg,0.141 mmol), COMU (60 mg, 0.141 mmol), DMF (3.0 mL) and DIPEA (0.223 mL,1.28 mmol). After stirring at RT for 20 min, the reaction mixture wasdiluted with EtOAc. The organic solution was washed with saturatedaqueous NaHCO₃ and brine, then dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude material was purified byreverse-phase HPLC to afford methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate as the bis-TFA salt (82.4 mg, 60%). MS (ESI) m/z 866 [M+H]⁺.¹H NMR (400 MHz, cd₃od) δ 7.94-7.67 (m, 4H), 7.59 (d, J=9.1 Hz, 1H),7.52 (s, 1H), 7.48-7.33 (m, 4H), 7.11 (d, J=18.7 Hz, 1H), 5.68 (d, J=6.3Hz, 1H), 5.48-5.33 (m, 1H), 5.23 (dd, J=24.1, 15.7 Hz, 1H), 5.17-5.03(m, 3H), 4.22 (dd, J=17.0, 9.6 Hz, 1H), 4.16-4.01 (m, 1H), 3.91 (d,J=24.1 Hz, 1H), 3.83-3.68 (m, 1H), 3.68-3.59 (m, 3H), 3.59-3.49 (m, 3H),3.38 (ddd, J=15.9, 9.6, 5.7 Hz, 2H), 3.28-3.14 (m, 5H), 3.10 (dd,J=14.0, 8.2 Hz, 1H), 3.00 (dd, J=17.8, 9.6 Hz, 1H), 2.92 (dd, J=14.5,6.7 Hz, 1H), 2.73-2.41 (m, 2H), 2.40-2.11 (m, 2H), 2.11-1.83 (m, 2H),1.54 (t, J=9.7 Hz, 2H), 1.24 (d, J=6.2 Hz, 1H), 1.06 (t, J=8.0 Hz, 1H),0.99 (d, J=6.8 Hz, 1H), 0.94 (d, J=6.6 Hz, 2H), 0.85 (d, J=6.7 Hz, 2H).

Example OI

tert-Butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylatewas prepared from(2S,4S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate by the samemethod employed in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,replacing (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid.

Methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

tert-Butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(137 mg, 0.179 mmol) was dissolved in DCM (5 mL) and HCl (4.0 M indioxane, 1 mL) was added. After stirring at RT for 1.5 h, the reactionmixture was concentrated under reduced pressure. The crude residue wastreated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (69 mg,0.39 mmol), HATU (149 mg, 0.393 mmol), DMF (2.0 mL) and DIPEA (0.31 mL,1.8 mmol). After stirring for 15 min at RT, the reaction mixture wasquenched with water and purified by HPLC to provide methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (123 mg). MS (ESI) m/z 880 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ8.48 (s, 1H), 8.05 (t, J=11.2 Hz, 1H), 7.92 (dd, J=19.7, 10.1 Hz, 2H),7.74 (s, 2H), 7.59-7.44 (m, 2H), 5.49 (s, 1H), 5.40 (dt, J=16.3, 8.1 Hz,1H), 5.31-5.15 (m, 3H), 4.47-4.10 (m, 4H), 3.86-3.44 (m, 12H), 3.39 (dd,J=13.2, 7.1 Hz, 6H), 2.94-2.57 (m, 4H), 2.25-1.94 (m, 4H), 1.02-0.82 (m,12H).

Example OJ

Methyl{(2S,3S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate was prepared from tert-Butyl(2S,4S)-2-(5-{2-[(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylateusing the same method employed in the synthesis of methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate, replacing with (S)-2-(methoxycarbonylamino)-3-methylbutanoicacid with (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid. MS(ESI) m/z 908 [M+H]⁺.

Example OK

tert-Butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatewas synthesized from(2S,4S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate using thesame methods described for the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,substituting(S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidine-2-carboxylicacid for(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid.

Methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate was synthesized from tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylateusing the same method employed for the synthesis of methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate substituting tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor tert-Butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 871 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 7.87 (ddd, J=20.5,15.3, 6.8 Hz, 4H), 7.65 (s, 1H), 7.50-7.38 (m, 5H), 7.17 (s, 1H), 5.41(d, J=24.5 Hz, 1H), 5.28 (t, J=8.3 Hz, 1H), 5.20 (d, J=7.3 Hz, 3H), 4.24(d, J=7.2 Hz, 1H), 4.12 (d, J=10.3 Hz, 1H), 4.03-3.94 (m, 1H), 3.89 (dd,J=15.4, 8.6 Hz, 1H), 3.77 (t, J=9.6 Hz, 1H), 3.72-3.64 (m, 4H),3.63-3.52 (m, 4H), 3.43 (qd, J=9.5, 5.6 Hz, 3H), 3.30 (s, 3H), 3.24-3.08(m, 2H), 2.97 (dd, J=11.6, 5.4 Hz, 2H), 2.59 (dt, J=21.1, 7.8 Hz, 3H),2.29 (s, 1H), 2.24-2.14 (m, 2H), 2.11-1.85 (m, 2H), 0.92 (dd, J=15.8,6.7 Hz, 6H).

Example OL

tert-Butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatewas prepared according to the method described for the synthesis oftert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,substituting tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor tert-Butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.

Methyl{(2S)-1-[(2S)-2-(9-{2-[(2S,4S)-1-[(2R)-2-amino-2-phenylacetyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas prepared according to the method described for the synthesis ofmethyl(S)-1-((2S,4S)-2-(2′-((2S,4S)-1-((R)-2-amino-2-phenylacetyl)-4-(methoxymethyl)pyrrolidin-2-yl)-1H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate,substituting methyl(S)-1-((2S,4S)-2-(2′-((2S,4S)-1-((R)-2-tert-butoxycarbonylamino-2-phenylacetyl)-4-(methoxymethyl)pyrrolidin-2-yl)-1H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamatewith

tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 811 [M+H]⁺.

Example OM

tert-Butyl (2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatewas synthesized from3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one, by thesame methods employed in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,substituting(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid for(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid and(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid for (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid.

tert-Butyl (2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatewas prepared according to the method described for the synthesis oftert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,substituting tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.

Methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate was prepared from tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylateaccording to the same method described for the synthesis of methyl(S)-1-((2S,4S)-2-(2′-((2S,4S)-1-((2S,3R)-2-methoxycarbonylamino-3-methoxybutanoyl)-4-(methoxymethyl)pyrrolidin-2-yl)-1H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate,substituting (2S,4S)-tert-Butyl2-(2′-((2S,4S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-methylpyrrolidin-2-yl)-1H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylatewith tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 866 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.44 (d, J=19.8 Hz,1H), 8.02 (t, J=8.6 Hz, 2H), 7.98-7.81 (m, 3H), 7.74 (dd, J=22.2, 13.6Hz, 2H), 7.63-7.41 (m, 2H), 5.79 (d, J=6.0 Hz, 1H), 5.42 (dt, J=43.3,21.5 Hz, 2H), 5.31-5.10 (m, 5H), 4.85-4.70 (m, 1H), 4.52 (d, J=3.8 Hz,1H), 4.31 (t, J=8.2 Hz, 1H), 4.17 (dd, J=20.8, 8.8 Hz, 1H), 3.80 (dt,J=19.0, 7.3 Hz, 2H), 3.73-3.63 (m, 7H), 3.63-3.49 (m, 3H), 3.39 (d,J=9.7 Hz, 4H), 3.35 (s, 5H), 3.28 (d, J=4.4 Hz, 3H), 2.84 (d, J=8.8 Hz,1H), 2.72 (dd, J=12.5, 6.6 Hz, 1H), 2.59-2.45 (m, 1H), 2.45-2.11 (m,4H), 2.11-1.82 (m, 2H), 1.56 (d, J=6.6 Hz, 3H), 1.35-1.21 (m, 1H),1.22-1.12 (m, 4H), 1.10-1.01 (m, 2H), 0.99 (d, J=6.6 Hz, 3H), 0.91 (d,J=6.7 Hz, 3H).

Example ON

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateMethyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate was prepared according to the method described for thesynthesis of methyl {(2S,3S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate substituting tert-butyl(2S,4S)-2-[9-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor tert-Butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 863 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.43 (d, J=24.6 Hz,1H), 8.01 (dt, J=16.1, 8.0 Hz, 1H), 7.95-7.78 (m, 2H), 7.77-7.64 (m,2H), 7.59-7.41 (m, 2H), 5.79 (d, J=5.8 Hz, 1H), 5.39 (dt, J=46.2, 23.1Hz, 1H), 5.27-5.07 (m, 3H), 4.85-4.72 (m, 1H), 4.42 (t, J=8.6 Hz, 1H),4.31 (d, J=7.9 Hz, 1H), 4.17 (dd, J=19.7, 8.7 Hz, 1H), 3.81 (dd, J=23.6,13.3 Hz, 1H), 3.69 (d, J=10.0 Hz, 5H), 3.60 (dd, J=14.7, 7.8 Hz, 2H),3.42 (s, 3H), 3.17 (d, J=6.1 Hz, 1H), 3.07 (s, 1H), 2.99-2.91 (m, 1H),2.85 (s, 1H), 2.73 (dd, J=12.5, 6.4 Hz, 1H), 2.62-2.48 (m, 1H),2.45-2.14 (m, 3H), 2.10-1.91 (m, 2H), 1.83 (s, 1H), 1.57 (d, J=6.6 Hz,3H), 1.44 (d, J=7.4 Hz, 1H), 1.34-1.23 (m, 1H), 1.20-0.96 (m, 5H), 0.90(dt, J=14.8, 6.7 Hz, 9H).

Example Oo

Methyl{(2S)-1-[(2S)-2-(5-{2-[(2S,4S)-1-[(2R)-2-amino-2-phenylacetyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate was prepared according to the method described for thesynthesis of methyl(S)-1-((2S,4S)-2-(2′-((2S,4S)-1-((R)-2-amino-2-phenylacetyl)-4-(methoxymethyl)pyrrolidin-2-yl)-1H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate,substituting tert-butyl(2S,4S)-2-[9-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor (2S,4S)-tert-butyl2-(2′-((2S,4S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-methylpyrrolidin-2-yl)-H,1′H-7,7′-binaphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 811 [M+H]⁺.

Example OP

Methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamatewas synthesized according to the protocol described for the preparationof methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate, substituting tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatefor tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,4,5,11tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 886 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.02-7.85 (m, 2H),7.85-7.68 (m, 2H), 7.58 (d, J=21.5 Hz, 1H), 7.55-7.35 (m, 4H), 7.31 (d,J=13.6 Hz, 1H), 5.43 (d, J=19.1 Hz, 1H), 5.28 (t, J=8.3 Hz, 1H),5.25-5.10 (m, 3H), 4.13 (t, J=9.5 Hz, 1H), 3.93-3.54 (m, 7H), 3.42 (qd,J=9.5, 5.5 Hz, 2H), 3.34 (d, J=7.9 Hz, 1H), 3.28 (s, 3H), 3.19 (t, J=7.8Hz, 2H), 3.00 (t, J=7.8 Hz, 2H), 2.74-2.46 (m, 3H), 2.44-2.15 (m, 2H),2.12-1.86 (m, 2H), 1.56 (d, J=6.7 Hz, 2H), 1.29 (d, J=6.3 Hz, 1H),1.15-1.01 (m, 1H), 0.98 (d, J=6.7 Hz, 2H), 0.88 (d, J=6.8 Hz, 2H).

Example OQ

(2S,4S)-1-Tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-ethoxypyrrolidine-1,2-dicarboxylate

To a slurry of 9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (2.50 g, 6.8 mmol) in MeCN (20 mL) was added(2S,4S)-1-(tert-butoxycarbonyl)-4-ethoxypyrrolidine-2-carboxylic acid(2.68 g, 10.3 mmol) and DIPEA (1.3 mL, 7.5 mmol). The reaction washeated with stirring to 50° C. for 18 h. The reaction was then cooled toroom temperature and diluted with EtOAc. The solution was washed withHCl (1N) and brine. The aqueous layers were backextracted with EtOAc andthe resulting organic layers were combined, dried (Na₂SO₄) andconcentrated under reduced pressure. The crude residue was purified bysilica column chromatography (15% to 50% EtOAc/Hexanes) to afford(2S,4S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-ethoxypyrrolidine-1,2-dicarboxylate (2.08 g, 56%).

Tert-butyl(2S,4S)-2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate

To a solution of (2S,4S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)4-ethoxypyrrolidine-1,2-dicarboxylate (2.08 g, 3.8 mmol) in a mixture oftoluene (30 mL) and methoxyethanol (4 mL) was added ammonium acetate(2.90 g, 37.7 mmol). The solution was heated with stirring to 80° C. for18 h. The reaction was then cooled to room temperature and diluted withEtOAc. The solution was washed with brine, and the resulting aqueouslayer was backextracted with EtOAc. The resulting organic layers werecombined, dried (Na₂SO₄), and concentrated under reduced pressure. Thecrude residue was purified by silica column chromatography (10% to 75%EtOAc (w/5% MeOH)/Hexanes) to afford tert-butyl(2S,4S)-2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate(0.99 g, 50%).

Tert-butyl(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate

To a solution of(2S,4S)-2-(9-chloro-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate(0.99 g, 1.9 mmol) in CH₂Cl₂ (18 mL) was added MnO₂ (4.52 g, 52.0 mmol).The resulting slurry was stirred at room temperature for 18 h. Thereaction was filtered through celite, washed with CH₂Cl₂, andconcentrated under reduced pressure. The crude residue was purified bysilica column chromatography (10% to 75% EtOAc (w/5% MeOH)/Hexanes) toafford tert-butyl(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate(0.71 g, 72%)

Methyl{(2S)-1-[(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of tert-butyl(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidine-1-carboxylate(0.46 g, 0.9 mmol) in a mixture of CH₂Cl₂ (9.0 mL) and MeOH (1.5 mL) wasadded HCl (in dioxanes, 4M, 6.5 mL, 26.0 mmol). The resulting solutionwas stirred at room temperature for 2 h. The solution was concentratedto dryness under reduced pressure. To the crude intermediate in CH₂Cl₂(10.0 mL) was added (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid(0.17 g, 0.9 mmol), HATU (0.41 g, 1.1 mmol), and DIPEA (0.5 mL, 2.9mmol). The resulting solution was stirred at room temperature for 48 hand diluted with CH₂Cl₂. The solution was washed with aqueous HCl (1N)and brine. The aqueous layers were backextracted with CH₂Cl₂ (2×). Theresulting organic layers were combined, dried (Na₂SO₄), and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (20% to 100% EtOAc (w/5% MeOH)/Hexanes to 80% MeOH/EtOAc)to afford methyl{(2S)-1-[(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (0.46 g, 90%).

Methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

To a solution of methyl{(2S)-1-[(2S,4S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-ethoxypyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.46 g, 0.84 mmol) in dioxane (8.5 mL) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (0.32 g, 1.3mmol), potassium acetate (0.25 g, 2.5 mmol),bis(dibenzylideneacetone)palladium (0.032 g, 0.035 mmol), and2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (Xphos, 0.032 g,0.067 mmol). The resulting solution was degassed with argon for 5 minand heated, with stirring, to 90° C. for 6 h. The reaction was cooled toroom temperature, diluted with EtOAc, and filtered through celite. Thecrude residue was purified by silica column chromatography (20% to 100%EtOAc (w/5% MeOH)/Hexanes to 90% MeOH/EtOAc) to afford methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate(0.41 g, 73%).

Tert-butyl(2S)-2-[5-(2-{(2S,4S)-4-ethoxy-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

To a solution of methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate(0.41 g, 0.61 mmol) in a mixture of DME (6.1 mL) and DMF (1.0 mL) wasadded (S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (0.39 g, 1.2mmol), tetrakis(triphenylphosphine)palladium (0.021 g, 0.018 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (0.030 g, 0.041mmol), and aqueous potassium carbonate (2M, 1.0 mL, 2.0 mmol). Thesolution was degassed with argon for 5 min and heated, with stirring, to85° C. for 6 h. The solution was cooled to room temperature and dilutedwith EtOAc. The organic layer was washed with water and brine. Theaqueous layers were backextracted with EtOAc (3×). The combined organiclayers were dried over Na₂SO₄ and concentrated under reduced pressure.The crude residue was purified by silica column chromatography (20% to100% EtOAc (w/5% MeOH)/Hexanes to 80% MeOH/EtOAc) to afford tert-butyl(2S)-2-[5-(2-{(2S,4S)-4-ethoxy-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(0.16 g, 33%).

Methyl{(2S)-1-[(2S)-2-(5-{2-[(2S,4S)-4-ethoxy-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of tert-butyl(2S)-2-[5-(2-{(2S,4S)-4-ethoxy-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(0.048 g, 0.062 mmol) in a mixture of CH₂Cl₂ (1.0 mL) and MeOH (0.25 mL)was added HCl (in dioxanes, 4M, 0.47 mL, 1.9 mmol). The solution wasstirred at room temperature for 3 h, and then concentrated to drynessunder reduced pressure. To the crude intermediate suspended in CH₂Cl₂(1.5 mL) was added (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid(0.012 g, 0.069 mmol), HATU (0.029 g, 0.076 mmol), and DIPEA (0.050 mL,0.28 mmol). The resulting solution was stirred at room temperature for1.5 h. The reaction was diluted with DMF and aqueous LiOH (2.5 M, 4drops) was added. The solution was concentrated to remove the CH₂Cl₂ andthe crude residue was purified by preparative reverse phase HPLC (10% to52% MeCN/water with 0.1% TFA). The desired fractions were combined andconcentrated under reduced pressure to remove volatile organics. Theaddition of aqueous sodium bicarbonate with stirring resulted inprecipitation of a white solid. The precipitate was filtered through amembrane filter and washed with water. Drying under reduced pressureafforded methyl{(2S)-1-[(2S)-2-(5-{2-[(2S,4S)-4-ethoxy-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (0.008 g, 17%). 1H-NMR: 400 MHz, (MeOD) δ: (Mixture ofrotamers) 8.37 (s, 1H), 7.97 (s, 2H), 7.37-7.76 (m, 5H), 5.38-5.54 (m,1H), 5.18 (s, 2H), 5.14-5.16 (m, 1H), 4.21-4.31 (m, 4H), 3.87-4.09 (m,1H), 3.79-3.85 (m, 2H), 3.66 (s, 3H), 3.64 (s, 3H), 3.46-3.55 (m, 2H),2.30-2.35 (m, 3H), 2.04-2.06 (m, 3H), 1.11 (m, 2H), 0.95 (d, 3H), 0.88(d, 3H). MS (ESI) m/z 836.02 [M+H]⁺.

Example OR

Methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-4-ethoxy-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl(2S)-2-[5-(2-{(2S,4S)-4-ethoxy-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(0.11 g, 0.14 mmol) in a mixture of CH₂Cl₂ (2.0 mL) and MeOH (0.5 mL)was added HCl (in dioxanes, 4M, 1.0 mL, 4.0 mmol). The solution wasstirred at room temperature for 3 h, and then concentrated to drynessunder reduced pressure. To the crude intermediate suspended in CH₂Cl₂(1.5 mL) was added (R)-2-(methoxycarbonylamino)-2-phenylacetic acid(0.044 g, 0.21 mmol) and DIPEA (0.075 mL, 0.43 mmol). The resultingsolution was cooled to −40° C. and COMU (0.096 g, 0.22 mmol) was added.The reaction was allowed to slowly warm to 0° C. over 1 h. The reactionwas diluted with DMF. The solution was concentrated to remove the CH₂Cl₂and the crude residue was purified by preparative reverse phase HPLC(10% to 55% MeCN/water with 0.1% TFA). The desired fractions werecombined and concentrated under reduced pressure to remove volatileorganics. The addition of aqueous sodium bicarbonate with stirringresulted in precipitation of a white solid. The precipitate was filteredthrough a membrane filter and washed with water. Drying under reducedpressure afforded methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-4-ethoxy-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.022 g, 18%). 1H-NMR: 400 MHz, (MeOD) δ: (Mixture ofrotamers) 8.28 (d, 1H), 7.88 (d, 1H), 7.52-7.70 (m, 3H), 7.28-7.38 (m,5H), 6.90-6.96 (m, 2H), 5.44-5.47 (m, 1H), 5.31 (s, 1H), 5.12 (s, 2H),4.16-4.48 (m, 3H), 3.81-3.19 (m, 1H), 3.62-3.76 (m, 2H), 3.58 (s, 3H),2.56 (s, 3H), 2.42-2.57 (m, 1H), 2.31 (m, 1H), 1.81-2.41 (m, 5H), 1.04(t, 3H), 0.87 (d, 3H), 0.81 (d, 3H). MS (ESI) m/z 869.55 [M+H]⁺.

Example OS

Methyl(2S,3R)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methoxy-1-oxobutan-2-ylcarbamate

To a solution of (S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (1.00 g, 3.2 mmol)in a mixture of CH₂Cl₂ (30 mL) and MeOH (5 mL) was added HCl (indioxane, 4 M, 11.5 mL, 46.0 mmol). The solution was stirred at 40° C.for 1h, cooled to room temperature, and concentrated to dryness underreduced pressure. To the crude intermediate suspended in CH₂Cl₂ (30 mL)was added (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (0.67g, 3.5 mmol), HATU (1.47 g, 3.8 mmol), and DIPEA (1.00 mL, 6.0 mmol).The resulting solution was stirred at room temperature for 24 h. DMF (2mL) and aqueous LiOH (2.5 M, 1 mL) were added and the reaction wasconcentrated to dryness under reduced pressure. The crude material wasdiluted with EtOAc and washed with H₂O and brine. The aqueous layerswere backextracted with EtOAc. The combined organic layers were driedover Na₂SO₄ and concentrated under reduced pressure. The crude residuewas purified by silica column chromatography (20% to 100% EtOAc (w/5%MeOH)/CH₂Cl₂) to afford methyl(2S,3R)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methoxy-1-oxobutan-2-ylcarbamate(1.2g, 100%).

Example OT

Tert-butyl(2S,4S)-2-[9-(2-{(2S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of tert-butyl4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]-naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(1.0 g, 3.2 mmol) in a mixture of DMSO (2.0 mL) and dioxanes (2.0 mL)was added methyl(2S,3R)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methoxy-1-oxobutan-2-ylcarbamate(0.24 g, 0.62 mmol), tetrakis(triphenylphosphine)palladium (0.050 g,0.043 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(0.030 g, 0.041 mmol), and aqueous potassium carbonate (2M, 0.65 mL, 1.3mmol). The solution was degassed with argon for 5 min and heated, withstirring, to 85° C. for 6 h. The solution was cooled to room temperatureand diluted with EtOAc. The organic layer was washed with water andbrine. The aqueous layers were backextracted with EtOAc (3×). Thecombined organic layers were dried over Na₂SO₄ and concentrated underreduced pressure. The crude residue was purified by silica columnchromatography (20% to 100% EtOAc (w/5% MeOH)/Hexanes to 60% MeOH/EtOAc)to afford tert-butyl(2S,4S)-2-[9-(2-{(2S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.20 g, 63%).

Methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl(2S,4S)-2-[9-(2-{(2S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]pyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.20 g, 0.26 mmol) in a mixture of CH₂Cl₂ (3.0 mL) and MeOH (0.5 mL)was added HCl (in dioxanes, 4M, 2.0 mL, 8.0 mmol). The solution wasstirred at 40° C. for 1 h, and then cooled to room temperature andconcentrated to dryness under reduced pressure. To the crudeintermediate suspended in CH₂Cl₂ (3.0 mL) was added(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (0.081 g, 0.39 mmol)and DIPEA (0.150 mL, 0.86 mmol). The resulting solution was cooled to−40° C. and COMU (0.180 g, 0.42 mmol) was added. The reaction wasallowed to slowly warm to room temperature over 30 min and maintainedfor 1.5 h. The solution was diluted with CH₂Cl₂ and washed with aqueousbicarbonate. The aqueous layer was backextracted with CH₂Cl₂. Thecombined organic layers were dried over Na₂SO₄ and concentrated underreduced pressure. The crude residue was purified by preparative reversephase HPLC (10% to 50% MeCN/water with 0.1% TFA). The desired fractionswere combined and concentrated under reduced pressure to remove volatileorganics. The addition of aqueous sodium bicarbonate with stirringresulted in precipitation of a white solid. The precipitate was filteredthrough a membrane filter and washed with water. Drying under reducedpressure afforded methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(0.10 g, 46%). 1H-NMR: 400 MHz, (MeOD) δ: (Mixture of rotamers) 8.34 (s,1H), 7.92-7.97 (m, 2H), 7.33-7.69 (m, 10H), 5.53 (s, 1H), 5.36-5.39 (m,1H), 5.15-5.21 (m, 3H), 4.44 (d, 1H), 3.86-3.93 (m, 2H), 3.68-3.75 (m,2H), 3.66 (s, 3H), 3.65 (s, 3H), 3.46-3.57 (m, 2H), 3.28 (s, 3H), 3.19(s, 3H), 2.47-2.60 (m, 3H), 2.22-2.36 (m, 4H), 1.99-2.08 (m, 3H), 1.15(d, 3H). MS (ESI) m/z 886.19 [M+H]⁺.

Example OU

Methyl(1-{4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methoxy-1-oxobutan-2-yl)carbamate

To a solution of tert-butyl4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(0.25 g, 0.41 mmol) in a mixture of CH₂Cl₂ (4.0 mL) and MeOH (1.0 mL)was added HCl (in dioxanes, 4M, 3.0 mL, 12.0 mmol). The resultingsolution was stirred at 40° C. for 45 min. The solution was cooled toroom temperature and concentrated to dryness under reduced pressure. Tothe crude intermediate in CH₂Cl₂ (4.0 mL) was added(2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (0.08 g, 0.42mmol), HATU (0.17 g, 0.45 mmol), and DIPEA (0.4 mL, 2.3 mmol). Theresulting solution was stirred at room temperature for 48 h and dilutedwith CH₂Cl₂. The solution was washed with brine. The aqueous layer wasbackextracted with CH₂Cl₂ (2×). The resulting organic layers werecombined, dried (Na₂SO₄), and concentrated under reduced pressure. Thecrude residue was purified by silica column chromatography (30% to 100%EtOAc (w/5% MeOH)/Hexanes to 80% MeOH/EtOAc) to afford methyl(1-{4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methoxy-1-oxobutan-2-yl)carbamate(0.24 g, 92%).

Tert-butyl(2S)-2-(5-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate

To a solution of methyl(1-{4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methoxy-1-oxobutan-2-yl)carbamate(0.15 g, 0.22 mmol) in a mixture of DMSO (2.0 mL) and dioxane (2.0 mL)was added (S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (0.15 g, 0.40mmol), tetrakis(triphenylphosphine)palladium (0.028 g, 0.024 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (0.018 g, 0.025mmol), and aqueous potassium carbonate (2M, 0.35 mL, 0.70 mmol). Thesolution was degassed with argon for 5 min and heated, with stirring, to90° C. for 6 h. The solution was cooled to room temperature and dilutedwith EtOAc. The organic layer was washed with water and brine. Theaqueous layers were backextracted with EtOAc (3×). The combined organiclayers were dried over Na₂SO₄ and concentrated under reduced pressure.The crude residue was purified by preparative reverse phase HPLC (10% to55% MeCN/water with 0.1% TFA). The desired fractions were combined andconcentrated under reduced pressure to remove volatile organics. Theremaining solution was basified with aqueous bicarbonate and extractedwith CH₂Cl₂ (3×). The combined organic layers were dried over Na₂SO₄ andconcentrated under reduced pressure to provide tert-butyl(2S)-2-(5-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate(0.013 g, 7%).

Methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methoxybutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl(2S)-2-(5-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate(0.013 g, 0.016 mmol) in a mixture of CH₂Cl₂ (0.5 mL) and MeOH (0.02 mE)was added HCl (in dioxanes, 4M, 0.20 mL, 0.80 mmol). The solution wasstirred at room temperature for 1 h, and then concentrated to drynessunder reduced pressure. To the crude intermediate suspended in CH₂Cl₂(0.5 mL) was added (R)-2-(methoxycarbonylamino)-2-phenylacetic acid(0.006 g, 0.029 mmol) and DIPEA (0.05 mL, 0.28 mmol). The resultingsolution was cooled to 0° C. and COMU (0.012 g, 0.028 mmol) was added.The reaction was stirred at 0° C. for 30 min. The solution was dilutedwith DMF and aqueous LiOH (2.5 M, 2 drops) and concentrated underreduced pressure to remove the CH₂Cl₂. The crude residue was purified bypreparative reverse phase HPLC (10% to 55% MeCN/water with 0.1% TFA).The desired fractions were combined and concentrated under reducedpressure to remove volatile organics. The addition of aqueous sodiumbicarbonate with stirring resulted in precipitation of a white solid.The precipitate was filtered through a membrane filter and washed withwater. Drying under reduced pressure afforded methyl{(1R)-2-[(2S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methoxybutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.008 g, 61%). ¹H-NMR: 400 MHz, (MeOD) δ: (Mixture ofrotamers) 8.37 (m, 1H), 7.96-7.98 (m, 2H), 7.60-7.79 (m, 3H), 7.35-7.52(m, 6H), 6.98-7.03 (m, 1H), 5.52 (s, 1H), 5.26-5.39 (m, 2H), 5.20 (s,2H), 4.44 (m, 1H), 4.27 (m, 1H), 3.64 (s, 6H), 3.50-3.57 (m, 3H), 3.37(s, 3H), 3.29-3.44 (m, 3H), 3.20 (s, 3H), 2.68-2.72 (m, 2H), 2.57-2.62(m, 2H), 1.89-2.15 (m, 6H), 1.18 (d, 3H). MS (ESI) m/z 885.73 [M+H]⁺.

Example OV

Methyl[(2S)-1-{(2S,4S)-3-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-2-azabicyclo[3.1.0]hex-2-yl}-3-methyl-1-oxobutan-2-yl]carbamate

Methyl[(2S)-1-{(2S,4S)-3-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-2-azabicyclo[3.1.0]hex-2-yl}-3-methyl-1-oxobutan-2-yl]carbamatewas prepared following the procedure for methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamateby substitution of(1S,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylicacid for(2S,4S)-1-(tert-butoxycarbonyl)-4-ethoxypyrrolidine-2-carboxylic acid.

Tert-butyl(2S,4S)-2-[5-(2-{(1S,3S,5S)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of methyl[(2S)-1-{(2S,4S)-3-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-2-azabicyclo[3.1.0]hex-2-yl}-3-methyl-1-oxobutan-2-yl]carbamate(0.19 g, 0.30 mmol) in a mixture of DMSO (2.0 mL) and dioxane (2.0 mL)was added (2S,4S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.20 g, 0.55 mmol), tetrakis(triphenylphosphine)palladium (0.035 g,0.030 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(0.025 g, 0.034 mmol), and aqueous potassium carbonate (2M, 0.5 mL, 1.0mmol). The solution was degassed with argon for 5 min and heated, withstirring, to 90° C. for 6 h. The solution was cooled to roomtemperature, diluted with EtOAc, and filtered through celite. Thefiltrate was concentrated under reduced pressure and purified by silicacolumn chromatography (2% to 25% CH₂Cl₂/MeOH) and preparative reversephase HPLC (10% to 55% MeCN/water with 0.1% TFA). The desired fractionswere combined and concentrated under reduced pressure to remove volatileorganics. The aqueous layer was basified with aqueous sodium bicarbonateand extracted with CH₂Cl₂ (3×). The organic layers were combine, driedover Na₂SO₄, and concentrated under reduced pressure to affordtert-butyl(2S,4S)-2-[5-(2-{(1S,3S,5S)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.025 g, 11%).

Methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(1S,3S,5S)-2-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl(2S,4S)-2-[5-(2-{(1S,3S,5S)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.025 g, 0.032 mmol) in a mixture of CH₂Cl₂ (1.0 mL) and MeOH (0.25 mL)was added HCl (in dioxanes, 4M, 0.50 mL, 2.0 mmol). The solution wasstirred at room temperature for 12 h, and then concentrated to drynessunder reduced pressure. To the crude intermediate suspended in CH₂Cl₂(0.5 mL) was added (R)-2-(methoxycarbonylamino)-2-phenylacetic acid(0.012 g, 0.057 mmol) and DIPEA (0.05 mL, 0.28 mmol). The resultingsolution was cooled to 0° C. and COMU (0.023 g, 0.054 mmol) was added.The reaction was stirred at 0° C. for 30 min. The solution was dilutedwith DMF and aqueous LiOH (2.5 M, 2 drops) and concentrated underreduced pressure to remove the CH₂Cl₂. The crude residue was purified bypreparative reverse phase HPLC (10% to 55% MeCN/water with 0.1% TFA).The desired fractions were combined and concentrated under reducedpressure to remove volatile organics. The addition of aqueous sodiumbicarbonate with stirring resulted in precipitation of a white solid.The precipitate was filtered through a membrane filter and washed withwater. Drying under reduced pressure afforded methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(1 S,3S,5S)-2-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.015 g, 55%). ¹H-NMR: 400 MHz, (MeOD) δ: (Mixture ofrotamers) 8.35 (m, 1H), 7.94-7.96 (m, 2H), 7.54-7.78 (m, 6H), 6.93-7.00(m, 1H), 5.72 (m, 1H), 5.46 (s, 1H), 5.19 (s, 2H), 5.14-5.16 (m, 1H),3.95 (m, 1H), 3.67 (s, 3H), 3.63 (s, 3H), 3.42-3.49 (m, 2H), 3.24 (s,3H), 2.67-2.78 (m, 2H), 2.41-2.62 (m, 3H), 2.01-2.13 (m, 2H), 1.86-1.99(m, 3H), 0.99-1.03 (m, 2H), 0.90 (d, 3H). MS (ESI) m/z 882.23 [M+H]⁺.

Example OW Tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

To a solution of methyl(1-{4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl)carbamate(0.47 g, 0.78 mmol) in a mixture of DMSO (4.0 mL) and dioxane (4.0 mL)was added methyl(2S,3S)-1-((2S,5S)-2-(5-iodo-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl)-3-methyl-1-oxopentan-2-ylcarbamate(0.26 g, 0.72 mmol), tetrakis(triphenylphosphine)palladium (0.090 g,0.078 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(0.061g, 0.083 mmol), and aqueous potassium carbonate (2M, 1.2 mL, 2.4mmol). The solution was degassed with argon for 5 min and heated, withstirring, to 90° C. for 6 h. The solution was cooled to roomtemperature, diluted with EtOAc, and filtered through celite. Thefiltrate was concentrated under reduced pressure and diluted with EtOAc.The organic solution was washed with water and brine and the aqueouslayers were backextracted with EtOAc. The combined organic layers weredried over Na₂SO₄ and concentrated under reduced pressure. The cruderesidue was purified by silica column chromatography (10% to 100% EtOAc(5% MeOH)/CH₂Cl₂) to afford tert-butyl(2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.25 g, 40%).

Methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl (2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.175 g, 0.21 mmol) in a mixture of CH₂Cl₂ (2.0 mL) and MeOH (0.5 mL)was added HCl (in dioxanes, 4M, 1.6 mL, 6.4 mmol). The solution wasstirred at 40° C. for 1 h, cooled to room temperature, and thenconcentrated to dryness under reduced pressure. To the crudeintermediate suspended in CH₂Cl₂ (3.0 mL) was added(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (0.070 g, 0.34 mmol)and DIPEA (0.15 mL, 0.86 mmol). The resulting solution was cooled to−40° C. and COMU (0.15 g, 0.35 mmol) was added. The reaction was warmedto room temperature over 30 min and diluted with CH₂Cl₂. The solutionwas washed with saturated aqueous sodium bicarbonate. The aqueous layerwas backextracted with CH₂Cl₂, and the combined organic layers weredried over Na₂SO₄ and concentrated under reduced pressure. The cruderesidue was purified by preparative reverse phase HPLC (10% to 58%MeCN/water with 0.1% TFA). The desired fractions were combined andconcentrated under reduced pressure to remove volatile organics. Theaddition of aqueous sodium bicarbonate with stirring resulted inprecipitation of a white solid. The precipitate was filtered through amembrane filter and washed with water. Drying under reduced pressureafforded methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(0.079 g, 41%). ¹H-NMR: 400 MHz, (MeOD) δ: (Mixture of rotamers) 8.36(m, 1H), 7.93-7.98 (m, 2H), 7.66-7.84 (m, 3H), 7.35-7.48 (m, 7H), 5.53(s, 1H), 5.36-5.39 (m, 1H), 5.17 (d, 2H), 5.08 (m, 1H), 4.14-4.35 (m,1H), 3.74 (m, 4H), 3.64 (s, 3H), 3.62 (s, 3H), 3.46 (m, 1H), 3.19 (s,3H), 2.76 (m, 1H), 2.46-2.60 (m, 3H), 2.24-2.35 (m, 1H), 2.08-2.18 (m,2H), 1.91 (m, 1H), 1.61-1.87 (m, 2H), 1.48 (d, 3H), 1.13-1.21 (m, 3H),0.80-0.97 (m, 3H). MS (ESI) m/z 898.24 [M+H]⁺.

Example OX

Methyl{(2S)-1-[(2S,4S)-2-(9-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of tert-butyl (2S,4S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.075 g, 0.09 mmol) in a mixture of CH₂Cl₂ (1.0 mL) and MeOH (0.25 mL)was added HCl (in dioxanes, 4M, 0.7 mL, 2.8 mmol). The solution wasstirred at 40° C. for 1 h, cooled to room temperature, and thenconcentrated to dryness under reduced pressure. To the crudeintermediate suspended in CH₂Cl₂ (3.0 mL) was added(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.020 g, 0.14 mmol),HATU (0.043 g, 0.11 mmol) and DIPEA (0.10 mL, 0.57 mmol). The reactionwas stirred at room temperature for 2 h. The reaction was diluted withDMF and aqueous LiOH (2.5 M, 3 drops) and the CH₂Cl₂ was removed underreduced pressure. The crude residue was purified by preparative reversephase HPLC (10% to 58% MeCN/water with 0.1% TFA). The desired fractionswere combined and concentrated under reduced pressure to remove volatileorganics. The addition of aqueous sodium bicarbonate with stirringresulted in precipitation of a white solid. The precipitate was filteredthrough a membrane filter and washed with water. Drying under reducedpressure afforded methyl{(2S)-1-[(2S,4S)-2-(9-{2-[(2S,5S)-1-{(2S,3S)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.031 g, 38%). ¹H-NMR: 400 MHz, (MeOD) δ: (Mixture of rotamers) 8.34(m, 1H), 7.91-9.97 (m, 2H), 7.50-7.81 (m, 3H), 7.35-7.38 (m, 2H),5.17-5.26 (m, 3H), 5.08 (m, 1H), 4.14-4.33 (m, 4H), 3.64 (s, 3H), 3.63(s, 3H), 3.51-3.59 (m, 3H), 3.37 (s, 3H), 2.71 (m, 1H), 2.55-2.59 (m,1H), 2.23-2.33 (m, 1H), 1.92-2.10 (m, 2H), 1.77-1.89 (m, 1H), 1.60 (m,1H), 1.48 (d, 1H), 1.11-1.22 (m, 2H), 0.81-0.98 (m, 12H). MS (ESI) m/z864.27 [M+H]⁺.

Example PF

tert-butyl(2S,4S)-4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate

The title compound was obtained as in example LQ but using(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid in place of (S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylicacid.

2-[4-(methoxymethyl)pyrrolidin-2-yl]-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazole

Tert-butyl(2S,4S)-4-(methoxymethyl)-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylate(310 mg, 0.507 mmol) was treated with 2 mL 1.25N HCl in ethanol andstirred at room temperature for 2h then at 50° C. for 2h. The reactionmixture was concentrated under reduced pressure to give a dark yellowsolid that was directly in the next step.

methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

A mixture of (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (107 mg,0.608 mmol), HATU (231 mg, 0.608 mmol) and 6 mL 10% DIPEA in DMF waspre-activated for 5 minutes, then it was added to the amine salt fromthe step above and allowed to stir overnight. The reaction mixture waspartitioned between ethyl acetate and saturated sodium bicarbonate. Theorganic phase was concentrated and purified by silica gelchromatography. (103 mg)

tert-butyl2-[5-(2-{1-[N-(methoxycarbonyl)valyl]-4-(methoxymethyl)pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

The title compound was obtained as in example LQ but using methyl[(2S)-1-{(2S,4S)-4-ethoxy-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin--yl}-3-methyl-1-oxobutan-2-yl]carbamate(103 mg, 0.154 mmol) in place of tert-butyl2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylateand methyl(S)-1-((2S,4S)-2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(58 mg, 0.154 mmol) in place of methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate.(50.0 mg)

methyl{1-[4-(methoxymethyl)-2-{9-[2-(4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl}pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

tert-butyl2-[5-(2-{1-[N-(methoxycarbonyl)valyl]-4-(methoxymethyl)pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(50 mg, 0.063 mmol) was treated with 2 mL 1.25N HCl in ethanol andheated at 60° C. for 2h, then it was concentrated under reduced pressureand pumped dry under high vacuum and used directly in the next step.

methyl{1-[2-{9-[2-(1-{[(methoxycarbonyl)amino](phenyl)acetyl}-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[,2-d]imidazol-2-yl}-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A mixture of (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (13 mg,0.063 mmol), COMU (30 mg, 0.069 mmol) in 0.500 mL DMF and DIPEA (0.033mL, 0.189 mmol) was allowed to preactivate for 15 minutes before it wasadded to the solid crude amine salt from the previous step and stirredovernight. The product was purified by reverse phase HPLC. The productwas converted to the free base by dissolution in 2 mL 1:1acetonitrile:methanol and passage through a prepacked cartridge ofpolymer supported carbonate. Concentration and drying gave an off whitepowder. (23.3 mg).

MS (ESI) m/z 883.8 [M+H]⁺

¹H NMR (CD₃CN) 8.176 (s, 1H), 7.778 (m, 1H), 7.596-7.521 (m, 4H),7.455-7.347 (m, 6H), 7.218 (s, 1H), 5.482 (s, 1H), 5.310 (m, 1H), 5.192(m, 1H), 4.999 (q, 2H, J=14 Hz), 4.372 (d, 1H, J=6.4 Hz), 4.279 (m, 1H),3.800-3.697 (m, 2H), 3.632 (s, 3H) 3.597-3.445 (m, 7H), 3.355 (s, 3H),2.876 (m, 2H), 2.761 (m, 1H), 2.583 (m, 2H), 2.220 (m, 2H), 1.764 (m,1H), 1.070 (d, 3H, J=6.4 Hz), 1.020 (d, 3H, J=6.4 Hz), 0.898 (d, 3H,J=6.4 Hz).

Example PG

tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,1-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

The title compound was obtained as in example LQ but using methyl[(2S)-3-methyl-1-{(2S,4S)-4-methyl-2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(307 mg, 0.481 mmol) in place of tert-butyl2-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidine-1-carboxylateand methyl(S)-1-((2S,4S)-2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate(181 mg, 0.481 mmol) in place of methyl(S)-1-((S)-2-(5-bromo-1H-imidazol-2-yl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-ylcarbamate.(200.8 mg)

methyl{(2S)-3-methyl-1-[(2S,4S)-4-methyl-2-(9-{2-[(2S,4S)-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(200 mg, 0.262 mmol) was treated with 2 mL 1.25N HCl in ethanol andheated at 60° C. for 2h, then it was concentrated under reduced pressureand pumped dry under high vacuum and used directly in the next step.

methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A mixture of (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (13 mg,0.063 mmol), COMU (30 mg, 0.069 mmol) in 1.5 mL DMF was allowed topreactivate for 5 minutes before it was added to a solution of the aminefrom the previous salt in 1.5 mL DMF and DIPEA (0.137 mL, 0.786 mmol)and stirred overnight. The product was purified by reverse phase HPLC.The product was converted to the free base by dissolution in 2 mL 1:1acetonitrile:methanol and passage through a prepacked cartridge ofpolymer supported carbonate. Concentration and drying gave an off whitepowder. (25.8 mg).

MS (ESI) m/z 853.8 [M+H]⁺.

¹H NMR (CD₃CN) 8.164 (s, 1H), 7.781 (m, 1H), 7.609 (m, 2H), 7.535 (m,2H), 7.433-7.305 (m, 6H), 7.229 (s, 1H), 5.482 (s, 1H), 5.290 (m, 1H),5.191 (m, 1H), 4.997 (m, 2H), 4.372 (d, 1H, J=6.4 Hz), 4.267 (m, 1H),3.735-3.445 (m, 10H), 2.573 (m, 4H), 2.197 (m, 2H), 2.017 (m, 1H), 1.760(m, 1H), 1.204 (d, 3H, J=6.4 Hz), 1.068 (d, 3H, J=6.4 Hz), 1.010 (d, 3H,J=6.8 Hz), 0.887 (d, 3H, J=6.8 Hz).

Example PH

tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[(benzyloxy)carbonyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

The title compound was obtained as in example OF (compound tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate)but using (S)-1-(benzyloxy carbonyl)pyrrolidine-2-carboxylic acid inplace of(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid.

tert-butyl(2S,4S)-4-(methoxymethyl)-2-(5-{2-[(2S)-pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate

A mixture of tert-butyl(2S,4S)-2-[5-(2-{(2S)-1-[(benzyloxy)carbonyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(724 mg, 0.96 mmol) and 70 mg 10% Pd/C in 20 mL ethanol was hydrogenatedat 1 atm overnight. Additional 10% Pd/C (300 mg) and a portion of solidNaHCO₃ was added and hydrogenation continued for 4 hours. Filtrationthrough celite and concentration of the filtrate under reduced pressuregave the product as a dark brown solid, 454 mg. Purification by reversephase HPLC gave 65 mg purified product.

methyl{(1R)-2-[(2S)-2-(9-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A mixture of (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (22 mg,0.105 mmol), COMU (45 mg, 0.069 mmol), and tert-butyl(2S,4S)-4-(methoxymethyl)-2-(5-{2-[(2S)-pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate(65 mg, 0.105 mmol) in 1.5 mL 10% DIPEA in DMF was stirred for 1.5h. Thereaction mixture was partitioned between ethyl acetate and saturatedsodium bicarbonate. The organic phase was dried over sodium sulphate,filtered and concentrated under reduced pressure. The crude intermediatewas treated with 8 mL 1.25N HCl in ethanol at 50° C. for 4h. Addedsaturated sodium bicarbonate and extracted the free base intodichloromethane. (106 mg).

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A mixture of methyl{(1R)-2-[(2S)-2-(9-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (55 mg, 0.077 mmol),(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (14 mg, 0.077 mmol),HATU (32 mg, 0.085 mmol) and 0.4 mL 10% DIPEA in DMF was stirred at roomtemperature for 1 hour. The product was purified by reverse phase HPLC.The product was converted to the free base by dissolution in 2 mL 1:1acetonitrile:methanol and passage through a prepacked cartridge ofpolymer supported carbonate. The eluent was concentrated, the taken upin 1% TFA in 1:1 acetonitrile:water, frozen, and lyophilized to give theproduct as a trifluoroacetate salt. (30.7 mg)

MS (ESI) m/z 869.9 [M+H]⁺.

methyl{(1R)-2-[(2S)-2-(9-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-threonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A mixture of methyl{(1R)-2-[(2S)-2-(9-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (51 mg, 0.072 mmol),(2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (14 mg, 0.072mmol), HATU (30 mg, 0.079 mmol) and 0.4 mL 10% DIPEA in DMF was stirredat room temperature for 1 hour. The product was purified by reversephase HPLC. The product was converted to the free base by dissolution in2 mL 1:1 acetonitrile:methanol and passage through a prepacked cartridgeof polymer supported carbonate. The eluent was concentrated, the takenup in 1% TFA in 1:1 acetonitrile:water, frozen, and lyophilized to givethe product as a trifluoroacetate salt. (24 mg)

MS (ESI) m/z 885.8 [M+H]+;

¹H NMR (CD₃CN) 7.635 (s, 1H), 7.434 (m, 3H), 7.330 (m, 4H), 7.233 (m,1H), 7.164 (m, 1H), 6.983 (m, 1H), 6.747 (m, 2H), 6.127 (m, 1H), 5.584(d, 1H, J=6.4 Hz), 5.431 (m, 1H), 5.145 (m, 1H), 4.729 (s, 2H), 4.442(m, 1H), 4.029 (m, 2H), 3.838 (m, 1H), 3.662-3.534 (m, 2H), 3.572 (s,3H) 3.552 (s, 3H), 3.444-3.310 (m, 3H), 3.240 (s, 3H), 3.225 (s, 3H),2.618 (m, 1H), 2.464 (m, 1H), 2.304 (m, 1H), 2.129 (m, 1H), 2.041 (m,1H), 1.899 (m, 2H), 1.107 (d, 3H, J=6.4 Hz).

Example PI

tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[(benzyloxy)carbonyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

The title compound was obtained as in example OF (compound tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate)but using (2S,5S)-1-(benzyloxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid in place of(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid.

tert-butyl(2S,4S)-4-(methoxymethyl)-2-(5-{2-[(2S,5S)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate

A mixture of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[(benzyloxy)carbonyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(830 mg, 1.08 mmol) and 100 mg 10% Pd/C in 20 mL ethanol washydrogenated at 1 atm overnight. Additional 10% Pd/C (300 mg) and aportion of solid NaHCO₃ was added and hydrogenation continued for 4hours. Filtration through celite and concentration of the filtrate underreduced pressure gave the product as a dark brown solid, 722 mg.Purification by reverse phase HPLC gave 100 mg purified product.

methyl{(2S,3R)-3-methoxy-1-[(2S,5S)-2-(9-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-1-oxobutan-2-yl}carbamate

A mixture of tert-butyl(2S,4S)-4-(methoxymethyl)-2-(5-{2-[(2S,5S)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidine-1-carboxylate(101 mg, 0.159 mmol), (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoicacid (30 mg, 0.159 mmol), HATU (61 mg, 0.159 mmol) and 2 mL 10% DIPEA inDMF was stirred at room temperature for 1.5 hours. Saturated sodiumbicarbonate was added and the product was extracted intodichloromethane, dried over sodium sulphate, filtered and concentratedunder reduced pressure. This crude product was treated with 5 mL 1.25NHCl in ethanol at 50° C. for 4h and then it was concentrated underreduced pressure. Saturated sodium bicarbonate was added and the productwas extracted into dichloromethane, dried over sodium sulphate, filteredand concentrated under reduced pressure. (74.6 mg)

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,5S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A mixture of methyl{(2S,3R)-3-methoxy-1-[(2S,5S)-2-(9-{2-[(2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-1-oxobutan-2-yl}carbamate (74.6 mg, 0.105 mmol),(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (18.5 mg, 0.105mmol), HATU (44 mg, 0.116 mmol) and 0.6 mL 10% DIPEA in DMF was stirredat room temperature for 1 hour. The product was purified by reversephase HPLC. (48.1 mg)

MS (ESI) m/z 866.1 [M+H]⁺.

Example PJ

tert-butyl(2S,4S)-2-(9-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-(trifluoromethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate

The title compound was prepared as in example OF for compound tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate,by using(2S,4S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylicacid in place of(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid and(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid in place of (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid.

methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(trifluoromethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

tert-butyl(2S,4S)-2-(9-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-4-(trifluoromethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylate(<0.412 mmol, crude from previous step) was treated with 6 mL 4N HCl indioxane at room temperature overnight and then at 50° C. for 1 hour.Diethyl ether (20 mL) was added and the precipitate of hydrochloridesalt was collected by vacuum filtration. (126 mg, 0.16 mmol). Thismaterial was combined with (R)-2-(methoxycarbonylamino)-2-phenylaceticacid (34 mg, 0.16 mmol), COMU (70 mg, 0.16 mmol), and 1.6 mL of 10%DIPEA in DMF. After 1 hour at room temperature, the mixture was addeddropwise into 25 mL saturated sodium bicarbonate, with stirring and theresulting precipitate was collected by vacuum filtration and washed with2 mL water. The product was purified, then re-purified by reverse phaseHPLC. (3.5 mg).

MS (ESI) m/z 938.1 [M+H]⁺.

Example PK

(2S,4S)-tert-butyl 2-formyl-4-methylpyrrolidine-1-carboxylate

A mixture of(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid(5.2g, 22.7 mmol), O,N-dimethylhydroxylamine hydrochloride (2.4g, 24.9mmol), HATU (10.4g, 27.2 mmol) and DIPEA (9.5 mL, 54.5 mmol) in 114 mLDMF was stirred at room temperature overnight. The mixture was extractedinto ethyl acetate and washed with saturated bicarbonate and water,dried over sodium sulphate, filtered, and concentrated. It was thendissolved in diethyl ether (100 mL) and washed with water to removeresidual DMF, dried, filtered, and concentrated to a pale yellow oil(5.30g, 19.5 mmol) of (2S,4S)-tert-butyl2-(methoxy(methyl)carbamoyl)-4-methylpyrrolidine-1-carboxylate.

(2S,4S)-tert-butyl2-(methoxy(methyl)carbamoyl)-4-methylpyrrolidine-1-carboxylate (5.30g,19.5 mmol) was dissolved in 120 mL THF, cooled to −78° C. and treatedwith lithium aluminum hydride (1M in THF, 19.5 mL, 19.5 mmol) dropwisevia addition funnel. After 1 hour, the mixture was brought to 0° C. andkept at that temperature for 2 hours. It was quenched by dropwiseaddition of a 50 mL solution of 3.0g KHSO4 in water, removed from theice bath, and stirred 15 minutes at room temperature. The product wasextracted with three 75 mL portions of ethyl acetate and washed withbrine. The organic phase was dried over sodium sulphate, filtered, andconcentrated to give crude (2S,4S)-tert-butyl2-formyl-4-methylpyrrolidine-1-carboxylate. (4.89g)

(2S,4S)-tert-butyl2-(1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

To a solution of (2S,4S)-tert-butyl2-formyl-4-methylpyrrolidine-1-carboxylate (4.89g, 22.9 mmol), ammoniumhydroxide (17 mL) and water (17 mL) was added, dropwise, glyoxal (40% inwater, 14.6 mL, 128 mmol) and the resulting mixture was stirred at roomtemperature overnight. Saturated sodium bicarbonate (100 mL) was addedand the mixture was extracted with four 75 mL portions ofdichloromethane. The organic phase was washed with water, dried oversodium sulphate, filtered and concentrated, and then purified by silicagel chromatography to give a total of 3.76g product.

(2S,4S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate

A mixture of (2S,4S)-tert-butyl2-(1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate (1.0g, 3.97mmol), iodine (2.22g, 8.75 mmol) and sodium carbonate (1.3g, 12.31 mmol)in 20 mL dioxane and 13.25 mL water was covered in foil and stirred atroom temperature overnight. The mixture was diluted with ethyl acetateand treated with 10% sodium thiosulfate (5 mL) and stirred for 10minutes. The organic phase was washed with brine, and then the aqueousphase was back extracted with ethyl acetate. The combined organic phaseswere dried over sodium sulphate, filtered and concentrated to providecrude (2S,4S)-tert-butyl2-(4,5-diiodo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(2.25g) as a pale yellow solid.

A solution of (2S,4S)-tert-butyl2-(4,5-diiodo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate(2.25g, 4.4 mmol) in 18 mL ethanol and 18 mL water was treated withsodium sulfite (5.59g, 44.4 mmol) and heated at 90° C. overnight. Themixture was partitioned between ethyl acetate and water. The aqueousphase was extracted with more ethyl acetate and the combined organicphase was washed with brine, dried over sodium sulphate, filtered,concentrated, and purified by silica gel chromatography to give 766 mg(2S,4S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate.

Example PL

(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid

To a solution of commercially available (2S,3aS,6aS)-benzyloctahydrocyclopenta[b]pyrrole-2-carboxylate hydrochloride (4.70 g, 16.68mmol) in methylene chloride (42 mL) was added Di-tert-butyl dicarbonate(7.28 g, 33.36 mmol), N,N-diisopropylethylamine (5.82 mL, 33.36 mmol)and 4-(Dimethylamino)pyridine (0.20 g, 1.67 mmol). The solution wasstirred under air for 16 hours. Upon completion, the reaction wasconcentrated in vacuo, diluted in ethyl acetate, and washed with 1N HCl.The aqueous layers were backextracted twice with ethyl acetate and thecombined organic layers were dried over sodium sulfate, filtered andconcentrated. The resulting residue was purified by silica gelchromatography (5-40% ethyl acetate in hexanes) to afford(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid which was used without further purification. MS (ESI) m/z 368.47[M+Na]⁺.

(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid

To a 250 mL round bottom flask charged with a stir bar and(2S,3aS,6aS)-2-benzyl 1-tert-butylhexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate (5.76 g, 16.68 mmol)was added 10% Palladium on carbon (1.77g). Ethanol was poured over themixture and the reaction mixture was evacuated and flushed with hydrogengas three times. The suspension was stirred at room temperature under anatmosphere of hydrogen for 24 hours. Upon completion, the reactionmixture was filtered through celite and concentrated to give (2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid (4.45g, >99%). MS (ESI) m/z 256.21 [M+H]+.

Example PM

tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

This compound was made in an analogous manner to tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylatesubstituting(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid for the initial alkylation of9-bromo-3-chloro-10,11-dihydro-6H-naphtho[2,3-c]chromen-8(9H)-one.Reactions in the synthesis of tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylategave similar product yields as in the synthesis of tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate.MS (ESI) m/z 774.1 [M+H]⁺.

Example PN

methyl{(1R)-2-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(0.128 g, 0.165 mmol) in a mixture of CH₂Cl₂ (1.6 mL) and MeOH (0.33 mL)was added HCl (4M in 1,4-dioxane, 1.24 mL, 4.9 mmol). The solution wasstirred at room temperature for 1.5 h and concentrated to dryness.

The intermediate was dissolved in CH₂Cl₂ (1.6 mL).(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (0.052 g, 0.25 mmol)and DIPEA (0.087 mL, 0.496 mmol) were then added to the solution. Thereaction mixture was cooled to −40° C. (external temperature,MeCN/CO₂(s) bath). COMU (0.113 g, 0.265 mmol) was then added andsolution was allowed to warm to 0° C. over 1.5 h. Upon completion byLCMS, the solution was diluted with DMF and concentrated. The crudeproduct was purified by preparative HPLC (Gemini column, 10-47% MeCN/H₂Owith 0.1% TFA) and the desired fractions were combined. The solution wasconcentrated until the aqueous layer remained and aqueous bicarbonate(sat.) was slowly added until the solution was basic. The resultingslurry was stirred at room temperature for 2h and filtered. Theresulting solid was dried in vacuo to provide methyl{(1R)-2-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate(0.068 g, 48%).

MS (ESI) m/z 865.7 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.44-8.30 (m, 1H),8.02-7.82 (m, 2H), 7.81-7.58 (m, 4H), 7.50-7.11 (m, 6H), 7.09-6.83 (m,2H), 5.72-5.45 (m, 2H), 5.41 (s, 1H), 5.34-5.28 (m, 1H), 5.22 (s, 3H),4.69-4.64 (m, 1H), 4.26-4.19 (m, 1H), 4.03-3.98 (m, 1H), 3.96-3.91 (m,1H), 3.66 (d, 4H), 2.98-2.91 (m, 1H), 2.88-2.83 (m, 1H), 2.58-2.48 (m,1H), 2.27-2.12 (m, 4H), 2.11-2.00 (m, 3H), 2.00-1.89 (m, 2H), 1.77-1.72(m, 1H), 1.31-1.04 (m, 3H), 0.93 (d, 6H).

Example PO

methyl{(2S)-1-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate(0.030 g, 0.039 mmol) in a mixture of CH₂Cl₂ (0.39 mL) and MeOH (0.078mL) was added HCl (4M in 1,4-dioxane, 0.29 mL, 1.16 mmol). The solutionwas stirred at room temperature for 1.5 h and concentrated to dryness.

The intermediate was dissolved in CH₂Cl₂ (0.39 mL).(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.007 g, 0.043 mmol)and DIPEA (0.020 mL, 0.116 mmol) were then added to the solution. HATU(0.018 g, 0.047 mmol) was added and solution was allowed to stir at roomtemp. Upon completion, the solution was diluted with DMF andconcentrated. The crude product was purified by preparative HPLC (Geminicolumn, 10-47% MeCN/H₂O with 0.1% TFA) and the desired fractions werecombined and lyophilized to provide methyl{(2S)-1-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.010g, 31%). MS (ESI) m/z 832.2 [M+H]⁺.

Example PP

methyl[(1S)-2-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl]carbamate

This compound was made in an analogous manner to methyl{(2S)-1-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate, substituting(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid for(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid to give methyl[(1S)-2-[2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl]carbamate(0.039 g, 56%). MS (ESI) m/z 874.34 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ8.58 (s, 2H), 8.26-8.08 (m, 2H), 7.96-7.75 (m, 4H), 7.65-7.54 (m, 5H),5.36-5.11 (m, 4H), 4.34-4.04 (m, 4H), 3.97-3.79 (m, 4H), 3.65 (s, 4H),3.53-3.44 (m, 2H), 2.68-2.47 (m, 4H), 2.32-2.02 (m, 7H), 1.95-1.82 (m,3H), 1.77-1.54 (m, 4H), 1.49-1.24 (m, 5H), 1.10-0.99 (m, 3H), 0.92-0.85(m, 4H).

Example PQ

tert-butyl2-[5-(2-{(2S,4S)-4-[(difluoromethoxy)methyl]-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate

This compound was made in an analogous manner to tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylatesubstituting(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid for the initial alkylation of9-bromo-3-chloro-10,11-dihydro-6H-naphtho[2,3-c]chromen-8(9H)-one.Reactions in the synthesis of tert-butyl2-[5-(2-{(2S,4S)-4-[(difluoromethoxy)methyl]-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylategave similar product yields as in the synthesis of tert-butyl2-[5-(2-{(2S,3aS,6aS)-1-[N-(methoxycarbonyl)-L-valyl]octahydrocyclopenta[b]pyrrol-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate.MS (ESI) m/z 815.04 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ 8.58 (s, 1H), 8.18(d, 1H), 7.96-7.85 (m, 3H), 7.70 (s, 1H), 7.60 (d, 1H), 7.50-7.38 (m,4H), 7.10 (s, 1H), 6.46 (t, 1H), 5.51 (s, 1H), 5.39-5.36 (m, 1H),5.31-5.28 (m, 2H), 4.43-4.36 (m, 1H), 4.24 (d, 1H), 4.13-4.02 (m, 3H),3.75-3.62 (m, 7H), 3.51-3.47 (m, 1H), 3.18-3.11 (m, 2H), 2.93-2.83 (m,2H), 2.75-2.69 (m, 1H), 2.47-2.36 (m, 2H), 2.23-2.09 (m, 3H), 2.01-1.94(m, 2H), 0.87 (dd, 6H).

Example PR

tert-butyl(2S,4S)-2-[5-(2-{(1R,3S,5R)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

This compound was made in an analogous manner to tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylatesubstituting(1R,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylicacid for the initial alkylation of9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one, andsubstituting (2S,4S)-tert-butyl2-(5-iodo-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidine-1-carboxylatefor the Suzuki-Miyara coupling. Reactions in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(1R,3S,5R)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylategavesimilar product yields as in the synthesis of tert-butyl(2R)-2-[5-(2-{(2S)-1-[N-(methoxycarbonyl)-L-valyl]pyrrolidin-2-yl}-3,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidine-1-carboxylate.MS (ESI) m/z 791.0 [M+H]⁺.

methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(1R,3S,5R)-2-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

To a solution of tert-butyl(2S,4S)-2-[5-(2-{(1R,3S,5R)-2-[N-(methoxycarbonyl)-L-valyl]-2-azabicyclo[3.1.0]hex-3-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.060 g, 0.076 mmol) in a mixture of CH₂Cl₂ (0.76 mL) and MeOH (0.15mL) was added HCl (4M in 1,4-dioxane, 0.570 mL, 2.28 mmol). The solutionwas stirred at room temperature for 2 h and concentrated to dryness.

The intermediate was dissolved in CH₂Cl₂ (0.76 mL).(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (0.024 g, 0.114 mmol)and DIPEA (0.040 mL, 0.228 mmol) were then added to the solution. Thereaction mixture was cooled to −40° C. (external temperature,MeCN/CO₂(s) bath). COMU (0.052 g, 0.122 mmol) was then added andsolution was allowed to warm to 0° C. over 1.5 h. Upon completion byLCMS, the solution was diluted with DMF and concentrated. The crudeproduct was purified by preparative HPLC (Gemini column, 10-45% MeCN/H₂Owith 0.1% TFA) and lyophilized to provide methyl{(1R)-2-[(2S,4S)-2-(5-{2-[(1R,3S,5R)-2-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-2-azabicyclo[3.1.0]hex-3-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.028 g, 42%). MS (ESI) m/z 881.8 [M+H]⁺. ¹H NMR (400 MHz,cd₃od) δ 8.45-8.33 (m, 1H), 8.02-7.94 (m, 1H), 7.91-7.75 (m, 2H),7.72-7.67 (m, 1H), 7.61 (s, 1H), 7.59-7.34 (m, 6H), 7.09-6.91 (m, 2H),5.62-5.38 (m, 2H), 5.29 (t, 1H), 5.24-5.09 (m, 3H), 4.61 (d, 1H),4.37-4.26 (m, 1H), 3.83-3.73 (m, 1H), 3.69-3.56 (m, 6H), 3.50-3.40 (m,1H), 3.20-3.11 (m, 1H), 2.99 (s, 1H), 2.83 (d, 1H), 2.63-2.50 (m, 2H),2.47-2.34 (m, 2H), 2.29-2.13 (m, 2H), 2.10-1.95 (m, 2H), 1.37-1.23 (m,3H), 1.19-1.10 (m, 1H), 1.03-0.78 (m, 7H).

Example PS

tert-butyl(2S,4S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-[(difluoromethoxy)methyl]pyrrolidine-1-carboxylate

This compound was made in an analogous manner to tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatesubstituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidfor the initial alkylation of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one, andsubstituting(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid for the other alkylation in the sequence. Reactions in thesynthesis of tert-butyl(2S,4S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-[(difluoromethoxy)methyl]pyrrolidine-1-carboxylategave similar product yields as in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 772.03 [M+H]⁺.

methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-4-[(difluoromethoxy)methyl]-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To a solution of tert-butyl(2S,4S)-2-(9-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-[(difluoromethoxy)methyl]pyrrolidine-1-carboxylate(0.081 g, 0.105 mmol) in a mixture of CH₂Cl₂ (1.05 mL) and MeOH (0.210mL) was added HCl (4M in 1,4-dioxane, 0.788 mL, 3.15 mmol). The solutionwas stirred at room temperature for 2 h and concentrated to dryness.

The intermediate was dissolved in CH₂Cl₂ (1.05 mL).(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.040 g, 0.231 mmol)and DIPEA (0.055 mL, 0.315 mmol) were then added to the solution. HATU(0.176 g, 0.462 mmol) was added and solution was allowed to stir at roomtemp. Upon completion, the solution was diluted with DMF andconcentrated. The crude product was purified by preparative HPLC (Geminicolumn, 10-45% MeCN/H₂O with 0.1% TFA) and the desired fractions werecombined. The solution was concentrated until the aqueous layer remainedand aqueous bicarbonate (sat.) was slowly added until the solution wasbasic. The resulting slurry was stirred at room temperature for 2h andfiltered. The resulting solid was dried in vacuo to provide methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-4-[(difluoromethoxy)methyl]-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (0.025 g, 27%). MS (ESI) m/z 886.1 [M+H]⁺. ¹H NMR (400 MHz,cd₃od) δ 8.49-8.25 (m, 2H), 8.08-7.82 (m, 2H), 7.79-7.27 (m, 5H), 6.45(t, 1H), 5.36-5.26 (m, 1H), 5.22-5.07 (m, 3H), 4.78-4.49 (m, 2H),4.45-4.19 (m, 3H), 4.16-4.05 (m, 2H), 3.99-3.92 (m, 1H), 3.85-3.71 (m,2H), 3.66 (s, 3H), 2.88-2.70 (m, 2H), 2.69-2.49 (m, 2H), 2.42-2.26 (m,2H), 2.23-2.10 (m, 2H), 2.07-1.87 (m, 3H), 1.51 (d, 2H), 1.34-1.20 (m,2H), 1.17-0.76 (m, 12H).

Example PT

methyl{(2S)-1-[(2S,4S)-4-[(difluoromethoxy)methyl]-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

This compound was made in an analogous manner to tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatesubstituting(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid for the initial alkylation of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one, andsubstituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidfor the other alkylation in the sequence. Reactions in the synthesis oftert-butyl(2S,4S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-[(difluoromethoxy)methyl]pyrrolidine-1-carboxylategave similar product yields as in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 772.31 [M+H]⁺.

methyl{(2S)-1-[(2S,4S)-4-[(difluoromethoxy)methyl]-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To tert-butyl(2S,4S)-2-(5-{2-[(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-[(difluoromethoxy)methyl]pyrrolidine-1-carboxylate(0.057 g, 0.074 mmol) in a mixture of CH₂Cl₂ (0.739 mL) and MeOH (0.148mL) was added HCl (4M in 1,4-dioxane, 0.555 mL, 2.218 mmol). Thesolution was stirred at room temperature for 2 h and concentrated todryness.

The intermediate was dissolved in CH₂Cl₂ (0.739 mL).(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.028 g, 0.163 mmol)and DIPEA (0.039 mL, 0.222 mmol) were then added to the solution. HATU(0.124 g, 0.325 mmol) was added and solution was allowed to stir at roomtemp. Upon completion, the solution was diluted with DMF andconcentrated. The crude product was purified by preparative HPLC (Geminicolumn, 10-46% MeCN/H₂O with 0.1% TFA) and the desired fractions werecombined and lyophilized to provide methyl{(2S)-1-[(2S,4S)-4-[(difluoromethoxy)methyl]-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.011 g, 17%). MS (ESI) m/z 886.1 [M+H]⁺. ¹H NMR (400 MHz, cd₃od) δ8.67-8.51 (m, 1H), 8.26-8.11 (m, 1H), 8.04-7.75 (m, 3H), 7.69-7.58 (m,2H), 6.43 (t, 1H), 5.41-5.15 (m, 4H), 4.48-3.90 (m, 6H), 3.82 (s, 1H),3.71-3.57 (m, 5H), 3.53-3.43 (m, 1H), 3.20-3.01 (m, 2H), 2.92-2.63 (m,3H), 2.60-2.25 (m, 4H), 2.15-1.86 (m, 4H), 1.57 (d, 3H), 1.24 (d, 2H),1.07 (dd, 2H), 0.98-0.77 (m, 9H).

Example PU

tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-(tert-butoxycarbonyl)-4-[(difluoromethoxy)methyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

This compound was made in an analogous manner to tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylatesubstituting(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid for the initial alkylation of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one, andsubstituting(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid for the other alkylation in the sequence. Reactions in thesynthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-(tert-butoxycarbonyl)-4-[(difluoromethoxy)methyl]pyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylategave similar product yields as in the synthesis of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate.MS (ESI) m/z 801.1 [M+H]⁺.

methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-4-[(difluoromethoxy)methyl]-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

To tert-butyl(2S,4S)-2-[5-(2-{(2S,4S)-1-(tert-butoxycarbonyl)-4-[(difluoromethoxy)methyl]pyrrolidin-2-yl}-1,11dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(0.092 g, 0.115 mmol) in a mixture of CH₂Cl₂ (1.15 mL) and MeOH (0.230mL) was added HCl (4M in 1,4-dioxane, 0.862 mL, 3.446 mmol). Thesolution was stirred at room temperature for 2 h and concentrated todryness.

The intermediate was dissolved in CH₂Cl₂ (1.149 mL).(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.044 g, 0.253 mmol)and DIPEA (0.060 mL, 0.345 mmol) were then added to the solution. HATU(0.192 g, 0.505 mmol) was added and solution was allowed to stir at roomtemp. Upon completion, the solution was diluted with DMF andconcentrated. The crude product was purified by preparative HPLC (Geminicolumn, 10-45% MeCN/H₂O with 0.1% TFA) and the desired fractions werecombined. The solution was concentrated until the aqueous layer remainedand aqueous bicarbonate (sat.) was slowly added until the solution wasbasic. The resulting slurry was stirred at room temperature for 2h andfiltered. The resulting solid was dried in vacuo to provide methyl{(2S)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-4-[(difluoromethoxy)methyl]-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (0.042 g, 40%). MS (ESI) m/z 916.30 [M+H]⁺. ¹H NMR (400 MHz,cd₃od) δ 8.55-8.25 (m, 1H), 8.15-7.85 (m, 2H), 7.83-7.26 (m, 5H), 6.44(t, 1H), 5.37-5.02 (m, 4H), 4.47-4.35 (m, 1H), 4.33-4.18 (m, 3H),4.15-3.90 (m, 3H), 3.81-3.45 (m, 11H), 3.39 (s, 3H), 2.90-2.27 (m, 5H),2.22-1.92 (m, 4H), 1.12-0.73 (m, 13H).

Example PX

(2S,4S)-1-tert-butyl 2-methyl4-((difluoromethoxy)methyl)pyrrolidine-1,2-dicarboxylate

A 100 mL round-bottom flask was charged with (2S,4S)-1-tert-butyl2-methyl 4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate (3.33 g, 12.84mmol), CuI (0.489 g, 2.56 mmol), and anhydrous acetonitrile (57.1 mL).The reaction was heated to 45° C. (ext. oil bath).2,2-difluoro-2-(fluorosulfonyl)acetic acid (2.655 mL, 25.68 mmol) wasadded at 45° C. over 30 minutes via syringe pump. The reaction washeated for 30 minutes. Upon completion as monitored by TLC, the reactionmixture was cooled to room temperature and concentrated in vacuo. Thecrude residue was diluted in EtOAc and washed with sodium bicarbonate(aq). The bicarbonate layer was back extracted with ethyl acetate twice.Combined organic layers were washed with brine, dried over sodiumsulphate, filtered and concentrated. The resulting residue was furtherpurified via silica gel chromatography (10 to 40% EtOAc/Hexanes) toafford (2S,4S)-1-tert-butyl 2-methyl4-((difluoromethoxy)methyl)pyrrolidine-1,2-dicarboxylate (2.41 g, 61%).MS (ESI) m/z 210.21 [M+H-Boc]⁺.

(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicAcid

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-((difluoromethoxy)methyl)pyrrolidine-1,2-dicarboxylate (2.41 g, 7.79mmol) in a mixture of THF (39 mL) and MeOH (15.6 mL) was added LiOH (2.5M aqueous, 15.6 mL, 38.9 mmol). The resulting solution was stirred atroom temperature for 1h. Upon completion by TLC the reaction mixture wasacidified with aqueous HCl (1N). The desired product was extracted withCH₂Cl₂ (3×). The combined organic layers were dried over Na₂SO₄ andconcentrated to provide(2S,4S)-1-(tert-butoxycarbonyl)-4-((difluoromethoxy)methyl)pyrrolidine-2-carboxylicacid (2.4 g, 99%). MS (ESI) m/z 294.96 [M−H]⁻. ¹H-NMR: 400 MHz,(acetone-d₆) δ (mixture of rotamers): 6.50 (t, 1H), 4.36-4.17 (m, 1H),3.93 (d, 2H), 3.77-3.67 (m, 1H), 3.63-3.59 (m, 1H), 3.26-3.12 (m, 1H),2.72-2.41 (m, 2H), 1.89-1.73 (m, 2H), 1.41 (s, 9H).

Example PY

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(150 mg, 0.19 mmol) in 1.25 N HCl in EtOH (3 mL) was stirred overnightthen heated to 50° C. for 3h. The reaction was concentrated and thecrude material dissolved in DMF (2 mL). To this solution was added asolution of (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (52 mg,0.25 mmol) and COMU (90 mg, 0.21 mmol). To the resulting solution wasadded diisopropylethylamine (0.099 mL, 0.57 mmol). After stirring for 2hat room temperature, the reaction was quenched with 1N HCl (0.200 mL)and purified by HPLC. After lyophilization, the TFA salt was dissolvedin EtOAc and washed with saturated NaHCO₃. The organic phase was driedover Na₂SO₄ and concentrated. The free base was then dissolved inMeCN/H₂O and lyophilized to afford methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(65 mg, 39%). LCMS-ESI⁺: calculated for C₄₉H₅₄N₈O₈: 882.4; observed[M+1]⁺: 884.1. Diagnostic peaks in NMR ¹H NMR (CD₃OD): 8.28 (s, 1H),8.21 (s, 1H), 8.04 (s, 1H), 7.91-7.01 (m, 10H), 3.62 (s, 3H), 3.34 (s,3H), 3.23 (s, 3H), 1.56 (d, 3H), 1.03 (d, 3H), 0.94 (d, 3H).

Example PY-1

9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

To3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(20.3g, 54.4 mmol) in DCM (365 mL) was added MeOH (22 mL) and pyridiniumtribromide (18.24 g, 57.0 mmol). After 2h, water was added (100 mL) andafter briefly agitating the layers split and the bottom organic layerwas collected. The organic layer was then washed with 1M HCl (100 mL)and the bottom organic layer containing9-bromo-3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-onewas collected. 400 MHz ¹H NMR (CDCl₃) 7.75 (d, J=8.1 Hz, 1H), 7.68 (s,1H), 7.61 (s, 1H), 7.42 (d, J=7.5 Hz, 1H), 7.24 (s, 1H), 5.13 (s, 2H),4.99-4.96 (m, 1H), 4.73 (dd, J=4.1, 4.1 Hz, 1H), 3.69-3.66 (m, 1H),3.58-3.53 (m, 1H), 3.35-3.27 (m, 1H), 2.96-2.90 (m, 1H), 2.58-2.44 (m,2H), C—OH not observed.

To9-bromo-3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(approx. 54.4 mmol) in DCM (365 mL) was added sodium bicarbonate (5.45g), sodium bromide (6.14 g), TEMPO (16.55 mg) and water (60 mL). Thesolution was cooled between 0-5° C. and 6% bleach (91.5 mL) was added.After 1h isopropyl alcohol (20 mL) was added and the reaction mixturewas warmed to room temperature. Agitation was stopped, the layersseparated and the lower organic layer was collected and concentratedremoving approximately 345 g of solvent. The slurry was filtered and thecake washed with 50 mL water and then 50 mL DCM (pre-cooled to 5° C.).The solids were collected and dried under vacuum to obtain9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(18.6 g, 76% yield). 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85(d, J=8.2 Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s,2H), 4.74 (dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H),2.99-2.92 (m, 1H), 2.59-2.46 (m, 2H); 100 MHz ¹³C NMR (CDCl₃) δ 190.4,189.6, 154.2, 136.6, 134.1, 133.9, 132.9, 131.8, 129.3, 127.2, 125.6,124.2, 123.3, 117.0, 68.1, 49.9, 31.8, 30.4, 25.5.

Example PY-2

9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A mixture of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (2.58g, 6.95 mmol), pyridinium tribromide (2.56 g, 8.0 mmol), dichloromethane(22 mL) and methanol (2.5 mL) was stirred at about 20° C. for 3 hours toobtain a slurry. The precipitated product was filtered, washed withdichloromethane (10 mL) and dried in a vacuum oven at 40° C. to give9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(2.62 g, 84% yield). 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85(d, J=8.2 Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s,2H), 4.74 (dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H),2.99-2.92 (m, 1H), 2.59-2.46 (m, 2H).

Example PY-3

3-((trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 300 mL flask equipped with an overhead stirrer and a reflux condenserunder an atmosphere of nitrogen was charged with3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (10.0g, 35.12mmol), powdered anhydrous tripotassium phosphate (22.4 g, 105.4 mmol),XPhos (1.34 g, 2.81 mmol), and PdCl₂(MeCN)₂ (364 mg, 1.40 mmol).Acetonitrile (140 mL) was added followed by TMSacetylene (18 mL, 141mmol). The mixture was heated to 65° C. After 6h, the reaction wasjudged complete, and the mixture was cooled to 20° C. The mixture wasfiltered through a fritted funnel, and the filtercake was washed withacetonitrile. The filtrate was concentrated to about 150 mL underreduced pressure and extracted with heptane (50 mL, 3×100 mL). N-Acetylcysteine (15 g) was added to the acetonitrile phase, and the mixture wasagitated for 5 h at 45° C. The mixture was cooled to ambienttemperature, filtered through a fritted funnel, and the filtercake waswashed with acetonitrile. The filtrate was concentrated to about 120 mLunder reduced pressure. Water (120 mL) was added and the mixture wasagitated for 40 minutes at 45° C. and then cooled to ambienttemperature. After 30 minutes the mixture was filtered through a frittedfunnel to provide3-((trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(4.07 g, 33.4% yield) as a yellow solid: 400 MHz ¹H NMR (CDCl₃) δ 7.65(d, J=8.1 Hz, 1H), 7.60 (s, 1H), 7.55 (s, 1H), 7.47 (dd, J=8.1, 1.4 Hz,1H), 7.27 (s, 1H), 5.06 (s, 2H), 2.95 (t, J=6.1 Hz, 2H), 2.67-2.59 (m,2H), 2.18-2.08 (m, 2H), 0.26 (s, 9H).

3-acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 20 mL vial with stir bar was charged with3-((trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(850 mg, 2.44 mmol) and TFA (9.8 mL). The solution was heated to 65° C.After 3 h, the reaction was judged complete. The mixture wasconcentrated under reduced pressure; the resulting residue was taken upin CH₂Cl₂ and loaded onto a prepacked 25g silica gel cartridge. Theproduct was purified by chromatography on a prepacked 80g silica gelcolumn eluting with a solvent gradient from 5% to 85% EtOAc/hexanes. Theproduct containing fractions were combined and concentrated to provide3-acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (616 mg, 86%):400 MHz ¹H NMR (CDCl₃) δ 8.00-7.94 (m, 1H), 7.81 (d, J=8.2 Hz, 1H), 7.77(s, 1H), 7.64 (s, 2H), 5.16 (s, 2H), 2.98 (t, J=6.1 Hz, 2H), 2.69-2.64(m, 2H), 2.63 (s, 3H), 2.21-2.09 (m, 2H).

9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 20 mL vial with a stir bar was charged with3-acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (100 mg, 0.366mmol), 9:1 CH₂Cl₂/MeOH (3.4 mL) and pyridinium tribromide (246 mg, 0.769mmol). The solution was heated to 35° C. After 30 minutes, the reactionwas judged complete. The mixture was cooled to ambient temperature,diluted with EtOAc (50 mL) and sequentially washed with saturatedaqueous Na₂S₂O₃ (20 mL), 2% aqueous NaHCO₃ (20 mL), water (20 mL), andbrine (10 mL). The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure resulting in9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(68 mg, 41%): 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85 (d, J=8.2Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s, 2H), 4.74(dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H), 2.99-2.92 (m,1H), 2.59-2.46 (m, 2H).

Example PY-4

(2S,5S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate

9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(1.43 g, 3.17 mmol) was treated with a solution of(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(800 mg, 3.49 mmol) in dichloromethane (14 mL) and K₂C03 (658 mg, 1.18mmol). The stirred reaction mixture was stirred at RT and diluted withCH₂Cl₂ and extracted 3×. The organic phase was washed with brine, thendried over MgSO₄, filtered and concentrated under reduced pressure toafford((2S,5S)-2-(2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl)1-tert-butyl 5-methylpyrrolidine-1,2-dicarboxylate (1.61 g, 84%).

This synthesis may be used to make a variety of compounds describedherein, including the compound exemplified in PY.

Example PZ

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(100 mg, 0.13 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (34mg, 0.20 mmol), HATU (54 mg, 0.14 mmol) and DMF (1.3 mL), thenN-methylmorpholine (0.043 mL, 0.39 mmol) was added dropwise. After 3h,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (91 mg, 82%).

LCMS-ESI⁺: calculated for C₄₆H₅₆N₈O₈: 848.4; observed [M+1]⁺: 850.2.

Example QA

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S,3S)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(119 mg, 0.15 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid(43 mg, 0.23 mmol), HATU (63 mg, 0.17 mmol) and DMF (2 mL), thenN-methylmorpholine (0.050 mL, 0.45 mmol) was added dropwise. After 3 hr,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S,3S)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(76 mg, 59%).

LCMS-ESI⁺: calculated for C₄₆H₅₆N₈O₉: 864.4; observed [M+1]⁺: 866.1.

Example QB

Methyl(2S,3S)-1-((2S,4S)-2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxopentan-2-ylcarbamate

(2S,4S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate (100 mg,0.13 mmol) in 1.25 N HCl in EtOH (15 mL) was heated to 50° C. for 3h andthen concentrated under reduced pressure. The crude residue was treatedwith (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid (625 mg,3.30 mmol), HATU (1.05 g, 2.77 mmol) and DMF (10 mL), then DIPEA (1.33mL, 7.62 mmol) was added dropwise. After 2h, the mixture was poured intosaturated aqueous NaHCO₃ and then extracted with EtOAc. The organicphase was washed successively with 5% aqueous LiCl and Brine. Theorganics were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(30 to 90% of 10% MeOH/EtoAc to Hexanes) afforded methyl(2S,3S)-1-((2S,4S)-2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxopentan-2-ylcarbamate(932 mg, 81%).

Tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate

(2S,4S)-Tert-butyl4-(methoxymethyl)-2-(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate(856 mg, 1.4 mmol), methyl(2S,3S)-1-((2S,4S)-2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl)-3-methyl-1-oxopentan-2-ylcarbamate(932 mg, 2.1 mmol), Pd(PPh₃)₄ (162 mg, 0.14 mmol), PdCl₂(dppf)₂ (102 mg,0.14 mmol), and K₂CO₃ (2M in H₂O, 2.31 mL, 4.62 mmol) were combined inDMSO (8 mL) and dioxanes (8 mL). The mixture was degassed with bubblingArgon for 10 min then heated to 95° C. for 1h. After cooling, thereaction mixture was diluted with EtOAc, and washed successively withsaturated aqueous NaHCO₃ and brine. The organics were dried over Na₂SO₄,filtered and concentrated under reduced pressure. The crude residue waspurified by silica column chromatography (1% to 20% MeOH/EtOAc) toafford tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(701 mg, 62%).

Methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate

A solution of tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(218 mg, 0.27 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h. The reaction was concentrated and the crude material dissolvedin DMF (3 mL). To this solution was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (73 mg, 0.35 mmol) andCOMU (127 mg, 0.30 mmol). To the resulting solution was addeddiisopropylethylamine (0.141 mL, 0.81 mmol). After stirring for 2h atroom temperature, the reaction was quenched with 1N HCl (0.200 mL) andpurified by HPLC. After lyophilization, the TFA salt was dissolved inEtOAc and washed with saturated NaHCO₃. The organic phase was dried overNa₂SO₄ and concentrated. The free base was then dissolved in MeCN/H₂Oand lyophilized to afford methyl{(1R)-2-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (121 mg, 50%). LCMS-ESI⁺: calculated for C₅₀H₅₆N₈O₈: 896.4;observed [M+1]⁺: 897.5.

Example QC

Methyl{(2S)-1-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(105 mg, 0.13 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (32mg, 0.18 mmol), HATU (59 mg, 0.16 mmol) and DMF (1.3 mL), thenN-methylmorpholine (0.043 mL, 0.39 mmol) was added dropwise. After 3h,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl{(2S)-1-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(80 mg, 71%).

LCMS-ESI⁺: calculated for C₄₇H₅₈N₈O₈: 862.4; observed [M+1]⁺: 864.2.

Example QD

Methyl{(2S,3R)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-allothreonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(105 mg, 0.13 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid(35 mg, 0.18 mmol), HATU (59 mg, 0.16 mmol) and DMF (1.3 mL), thenN-methylmorpholine (0.043 mL, 0.39 mmol) was added dropwise. After 3 hr,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl{(2S,3R)-1-[(2S,4S)-2-(5-{2-[(2S,4S)-1-[N-(methoxycarbonyl)-O-methyl-L-allothreonyl]-4-(methoxymethyl)pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-4-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate (92 mg, 81%).

LCMS-ESI: calculated for C₄₇H₅₈N₈O₉: 878.4; observed [M+1]⁺: 879.3.

Example QE

Methyl{(3R)-1-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

tert-butyl(2S,4S)-2-[9-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-alloisoleucyl]-4-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-4-(methoxymethyl)pyrrolidine-1-carboxylate(105 mg, 0.13 mmol) in 1.25 N HCl in EtOH (3 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid(34 mg, 0.18 mmol), HATU (59 mg, 0.16 mmol) and DMF (1.3 mL), thenN-methylmorpholine (0.043 mL, 0.39 mmol) was added dropwise. After 3h,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl{(3R)-1-[(2S,4S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-2-[(methoxycarbonyl)amino]-3-methylpentanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate(98 mg, 86%).

LCMS-ESI⁺: calculated for C₄₈H₆₀N₈O₈: 876.5; observed [M+1]⁺: 878.2.

Example QF

(2S,5S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate

To a solution of9-bromo-3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (1.41 g,3.88 mmol) in MeCN (17 mL) was added(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(980 mg, 4.27 mmol) and DIPEA (1.49 mL, 8.54 mmol). After stirring for18 h at 50° C., the solution was diluted with EtOAc and washedsuccessively with 1N HCl, saturated aqueous NaHCO₃ and brine. Theorganics were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(10% to 30% EtOAc/hexanes) to afford (2S,5S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate (1.63 g, 81%).

(2S,5S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-(methyl)pyrrolidine-1-carboxylate

(2S,5S)-1-tert-butyl2-(3-chloro-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate (1.63 g, 3.18 mmol) was addedtoluene (30 mL), 2-methoxyethanol (3 mL), and ammonium acetate (3.68 g,77.1 mmol) and the solution was heated to reflux overnight. Thefollowing morning, the solution was cooled to rt and was diluted withEtOAc and washed successively with water, saturated aqueous NaHCO₃ andbrine. The organics were dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The crude residue was purified by silica columnchromatography (40% to 80% EtOAc/hexanes) to afford (2S,5S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(1.13 g, 72%).

((2S,5S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate

To a solution of (2S,5S)-tert-butyl2-(9-chloro-4,5-dihydro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-(methyl)pyrrolidine-1-carboxylate(1.13 g, 2.3 mmol) in CH₂Cl₂ (25 mL) was added MnO₂ (9.98 g, 115 mmol).The reaction mixture was stirred overnight then filtered over celite.The filter cake was washed with copious CH₂Cl₂ and MeOH, and thefiltrate was concentrated under reduced pressure to afford the crudeproduct (2S,5S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(931 mg, 83%).

Methyl{(2S)-1-[(2S,5S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

(2S,5S)-tert-butyl2-(9-chloro-5H-naphtho[c,g]chromeno[8,9-d]imidazol-2-yl)-5-methylpyrrolidine-1-carboxylate(931 mg, 1.9 mmol) in 1.25 N HCl in EtOH (8 mL) was heated to 50° C. for3h and then concentrated under reduced pressure. The crude residue wastreated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (499 mg,2.9 mmol), HATU (795 mg, 2.1 mmol) and DMF (10 mL), thenN-methylmorpholine (0.627 mL, 5.7 mmol) was added dropwise. Afterstirring for 1 h, the reaction was diluted with EtOAc and washedsuccessively with saturated aqueous NaHCO₃, 5% LiCl, and brine. Theorganics were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude residue was purified by silica column chromatography(50% to 100% EtOAc/hexanes) to afford methyl{(2S)-1-[(2S,5S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (950 mg, 91%).

Methyl[(2S)-3-methyl-1-{(2S,5S)-2-methyl-5-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate

To methyl{(2S)-1-[(2S,5S)-2-(9-chloro-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(950 mg, 1.74 mmol) in dioxane (17 mL) was added bis(pinacolato)diboron(662 mg, 2.61 mmol), KOAc (512 mg, 5.22 mmol), X-Phos (25 mg, 0.05mmol), and Pd₂dba₃ (80 mg, 0.08 mmol). The solution was degassed with N2for 10 min, then heated to 90° C. for 16 h. The solution was cooled tort, diluted with EtOAc, washed with saturated aqueous NaHCO₃, brine,dried with Na₂SO₄, and concentrated. Purification by silica gelchromatography (30% to 75% gradient using 5% MeOH/EtOAc to Hexanes) toafford methyl[(2S)-3-methyl-1-{(2S,5S)-2-methyl-5-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(800 mg, 72%).

tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate

To a solution of[(2S)-3-methyl-1-{(2S,5S)-2-methyl-5-[9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-1-oxobutan-2-yl]carbamate(269 mg, 0.42 mmol), (2S,4S)-tert-butyl2-(5-bromo-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate (206 mg,0.54 mmol), tetrakis(triphenylphosphine) palladium(0) (49 mg, 0.042mmol) and dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)(31 mg, 0.042 mmol) in DMSO (3 mL) and dioxanes (3 mL) was added asolution of potassium carbonate (2M in water, 0.69 mL, 1.39 mmol). Theresulting mixture was degassed and then heated to 95° C. for 2h. Aftercooling to room temperature, the reaction was diluted with ethylacetate. The organics were washed with saturated sodium bicarbonate andbrine, dried over Na₂SO₄ and concentrated. The crude residue waspurified by flash chromatography (1 to 20% MeOH/EtOAc) to yieldtert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(202 mg, 63%).

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

A solution of tert-butyl(2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(80 mg, 0.11 mmol) in 1.25 N HCl in EtOH (2 mL) was heated to 50° C. for3h. The reaction was concentrated and the crude material dissolved inDMF (1.5 mL). To this solution was added a solution of(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (29 mg, 0.14 mmol) andCOMU (52 mg, 0.12 mmol). To the resulting solution was addeddiisopropylethylamine (0.057 mL, 0.33 mmol). After stirring for 2h atroom temperature, the reaction was quenched with 1N HCl (0.200 mL) andpurified by HPLC. After lyophilization, the TFA salt was dissolved inEtOAc and washed with saturated NaHCO₃. The organic phase was dried overNa₂SO₄ and concentrated. The free base was then dissolved in MeCN/H₂Oand lyophilized to afford methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate:(42 mg, 45%). LCMS-ESI⁺: calculated for C₄₈H₅₂N₈O₇: 852.4; observed[M+1]⁺: 854.2.

Example QG

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

tert-butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(60 mg, 0.079 mmol) in 1.25 N HCl in EtOH (2 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (21mg, 0.12 mmol), HATU (36 mg, 0.095 mmol) and DMF (1.5 mL), thenN-methylmorpholine (0.027 mL, 0.24 mmol) was added dropwise. After 3h,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(33 mg, 51%).

LCMS-ESI⁺: calculated for C₄₅H₅₄N₈O₇: 818.4; observed [M+1]⁺: 820.2.

Example QH

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S,3R)-3-methoxy-2-[(methoxycarbonyl)amino]butanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

tert-butyl (2S,4S)-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-4-methylpyrrolidine-1-carboxylate(20 mg, 0.079 mmol) in 1.25 N HCl in EtOH (2 mL) was heated to 50° C.for 3h and then concentrated under reduced pressure. The crude residuewas treated with (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid(8 mg, 0.04 mmol), HATU (12 mg, 0.03 mmol) and DMF (0.5 mL), thenN-methylmorpholine (0.009 mL, 0.078 mmol) was added dropwise. After 3h,the mixture was quenched with 1N HCl (0.100 mL) and then purified byHPLC to afford methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-4-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(7.5 mg, 35%).

LCMS-ESI⁺: calculated for C₄₅H₅₄N₈O₈: 834.4; observed [M+1]⁺: 835.7.

Compounds 478-647

Using procedures similar to those described herein, the followingcompounds of the disclosure were prepared.

LCMS (observed # Compound (M + H)⁺) 478

879.4  493

838.2  494

837.3  495

835.73 498

835.34 499

823.35 503

837.35 507

865.32 510

880.0  516

836.04 518

803.2  526

806.11 536

838.29 588

842.10 602

847.99 603

822.02 617

854.19 626

885.80 643

878.15 646

881.66

Biological Assays

Effect of Serum Proteins on Replicon Potency:

Replicon assays are conducted in normal cell culture medium (DMEM+10%FBS) supplemented with physiologic concentrations of human serum albumin(40 mg/mL) or α-acid glycoprotein (1 mg/mL). EC_(50S) in the presence ofhuman serum proteins are compared to the EC₅₀ in normal medium todetermine the fold shift in potency.

MT-4 Cell Cytotoxicity:

MT4 cells are treated with serial dilutions of compounds for a five dayperiod. Cell viability is measured at the end of the treatment periodusing the Promega CellTiter-Glo assay and non-linear regression isperformed to calculate CC₅₀.

Compound Concentration Associated with Cells at EC₅₀:

Huh-luc cultures are incubated with compound at concentrations equal toEC₅₀. At multiple time points (0-72 hours), cells are washed 2× withcold medium and extracted with 85% acetonitrile; a sample of the mediaat each time-point will also be extracted. Cell and media extracts areanalyzed by LC/MS/MS to determine the Molar concentration of compoundsin each fraction. Representative compounds of the disclosure have shownactivity.

Solubility and Stability:

Solubility is determined by taking an aliquot of 10 mM DMSO stocksolution and preparing the compound at a final concentration of 100 μMin the test media solutions (PBS, pH 7.4 and 0.1 N HCl, pH 1.5) with atotal DMSO concentration of 1%. The test media solutions are incubatedat room temperature with shaking for 1 hr. The solutions will then becentrifuged and the recovered supernatants are assayed on the HPLC/UV.Solubility will be calculated by comparing the amount of compounddetected in the defined test solution compared to the amount detected inDMSO at the same concentration. Stability of compounds after an 1 hourincubation with PBS at 37° C. will also be determined.

Stability in Cryopreserved Human, Dog, and Rat Hepatocytes:

Each compound is incubated for up to 1 hour in hepatocyte suspensions(100 μl, 80,000° Cells per well) at 37° C. Cryopreserved hepatocytes arereconstituted in the serum-free incubation medium. The suspension istransferred into 96-well plates (50 μL/well). The compounds are dilutedto 2 μM in incubation medium and then are added to hepatocytesuspensions to start the incubation. Samples are taken at 0, 10, 30 and60 minutes after the start of incubation and reaction will be quenchedwith a mixture consisting of 0.3% formic acid in 90% acetonitrile/10%water. The concentration of the compound in each sample is analyzedusing LC/MS/MS. The disappearance half-life of the compound inhepatocyte suspension is determined by fitting the concentration-timedata with a monophasic exponential equation. The data will also bescaled up to represent intrinsic hepatic clearance and/or total hepaticclearance.

Stability in Hepatic S9 Fraction from Human, Dog, and Rat:

Each compound is incubated for up to 1 hour in S9 suspension (500 μl, 3mg protein/mL) at 37° C. (n=3). The compounds are added to the S9suspension to start the incubation. Samples are taken at 0, 10, 30, and60 minutes after the start of incubation. The concentration of thecompound in each sample is analyzed using LC/MS/MS. The disappearancehalf-life of the compound in S9 suspension is determined by fitting theconcentration-time data with a monophasic exponential equation.

Caco-2 Permeability:

Compounds are assayed via a contract service (Absorption Systems, Exton,Pa.). Compounds are provided to the contractor in a blinded manner. Bothforward (A-to-B) and reverse (B-to-A) permeability will be measured.Caco-2 monolayers are grown to confluence on collagen-coated,microporous, polycarbonate membranes in 12-well Costar TRANSWELL®plates. The compounds are dosed on the apical side for forwardpermeability (A-to-B), and are dosed on the basolateral side for reversepermeability (B-to-A). The cells are incubated at 37° C. with 5% CO₂ ina humidified incubator. At the beginning of incubation and at 1 hr and 2hr after incubation, a 200-μL aliquot is taken from the receiver chamberand replaced with fresh assay buffer. The concentration of the compoundin each sample is determined with LC/MS/MS. The apparent permeability,Papp, is calculated.

Plasma Protein Binding:

Plasma protein binding is measured by equilibrium dialysis. Eachcompound is spiked into blank plasma at a final concentration of 2 μM.The spiked plasma and phosphate buffer is placed into opposite sides ofthe assembled dialysis cells, which will then be rotated slowly in a 37°C. water bath. At the end of the incubation, the concentration of thecompound in plasma and phosphate buffer is determined. The percentunbound is calculated using the following equation:

${\% \mspace{14mu} {Unbound}} = {100 \cdot \left( \frac{C_{f}}{C_{b} + C_{f}} \right)}$

Where C_(f) and C_(b) are free and bound concentrations determined asthe post-dialysis buffer and plasma concentrations, respectively.

CYP450 Profiling:

Each compound is incubated with each of 5 recombinant human CYP450enzymes, including CYP1A2, CYP2C9, CYP3A4, CYP2D6 and CYP2C19 in thepresence and absence of NADPH. Serial samples will be taken from theincubation mixture at the beginning of the incubation and at 5, 15, 30,45 and 60 minutes after the start of the incubation. The concentrationof the compound in the incubation mixture is determined by LC/MS/MS. Thepercentage of the compound remaining after incubation at each time pointis calculated by comparing with the sampling at the start of incubation.

Stability in Rat, Dog, Monkey and Human Plasma:

Compounds will be incubated for up to 2 hours in plasma (rat, dog,monkey, or human) at 37° C. Compounds are added to the plasma at finalconcentrations of 1 and 10 μg/mL. Aliquots are taken at 0, 5, 15, 30,60, and 120 minutes after adding the compound. Concentration ofcompounds and major metabolites at each time point are measured byLC/MS/MS.

Evaluation of Cell-Based Anti-HCV Activity:

Antiviral potency (EC₅₀) was determined using a Renilla luciferase(RLuc)-based HCV replicon reporter assay. To perform the assay forgenotype 1 and 2a JFH-1, stable HCV 1a RLuc replicon cells (harboring adicistronic genotype 1a H77 replicon that encodes a RLuc reporter),stable HCV 1b RLuc replicon cells (harboring a dicistronic genotype 1bCon1 replicon that encodes a RLuc reporter), or stable HCV 2a JFH-1 Rlucreplicon cells (harboring a dicistronic genotype 2a JFH-1 replicon thatencodes a RLuc reporter; with L31 present in NSSA) were dispensed into384-well plates for EC₅₀ assays. To perform the assay for genotype 2a(with M31 present in NSSA) or 2b, NS5A chimeric genotype 2a JFH-1replicons that encodes a RLuc-Neo reporter and either genotype 2a J6strain NS5A gene or genotype 2b MD2b-1 strain NSSA gene (both with M31present) respectively, were either transiently transfected (t) intoHuh-Lunet cells or were established as stably replicating replicon cells(s) is provided. Either cells were dispensed into 384-well plates forEC₅₀ assays. To perform the assay for genotype 3 and 4, NS5A chimericgenotype 1b Con1 replicons that encodes a Pi-RLuc reporter and eithergenotype 3a S52 strain NS5A gene or genotype 4a ED43 strain NS5A generespectively, were transiently transfected (t) into Huh-Lunet cells,which were subsequently dispensed into 384-well plates. Compounds weredissolved in DMSO at a concentration of 10 mM and diluted in DMSO eithermanually or using an automated pipeting instrument. Serially 3-folddiluted compounds were either manually mixed with cell culture media andadded to the seeded cells or directly added to the cells using anautomated instrument. DMSO was used as a negative (solvent; noinhibition) control, and the protease inhibitor ITMN-191 was included ata concentration >100×EC₅₀ as a positive control. 72 hours later, cellswere lysed and Renilla luciferase activity quantified as recommended bythe manufacturer (Promega-Madison, Wis.). Non-linear regression wasperformed to calculate EC₅₀ values.

To determine the antiviral potency (EC₅₀) against resistance mutants,resistance mutations, including M28T, Q30R, Q30H, L31M, and Y93C ingenotype 1a NS5A and Y93H in genotype 1b NSSA, were introducedindividually into either 1a Pi-Rluc or 1b Pi-Rluc replicons by sitedirected mutagenesis. Replicon RNA of each resistant mutant wastransiently transfected into Huh-7-derived cured-51 cells and antiviralpotency was determined on these transfected cells as described above.

The EC₅₀ ranges for genotype 1a, 1a Q30R, and 2a JFH are as follows:A≥44 nM, B=1 nM to 43.99 nM, C<1 nM. The EC₅₀ ranges for genotype 2a J6,2b, 3a, and 4a are as follows: A≥5 nM, B=1 nM to 4.99 nM, C<1 nM. TheEC₅₀ ranges for genotype 2a J6, 2b, and 4a correspond to the assay oftransiently transfected cells (t). If this data is unavailable, the EC₅₀range for the stably replicating cells (s) is provided.

IV and PO Single Dose Pharmacokinetic Studies in SD Rats:

The pharmacokinetics of selected compounds was characterized in maleSprague-Dawley (SD) rats (250-300g). In this study, two groups of navepurebred SD rats (N=3 per group, fasted over night) received theselected compound either as an intravenous (IV) infusion (1 mg/kg over30 minutes) via the jugular vein or by oral gavage (2 mg/kg). Theintravenous (IV) dosing vehicle was 5% ethanol, 35% polyethylene glycol400 (PEG 400) and 60% water pH 2.0. The oral dosing vehicle was 5%ethanol, 55% PEG 400 and 40% citrate buffer pH 2.2.

Serial blood samples (approximately 0.3 mL each) were collected fromjugular vein or other suitable vein at specified time points. For the IVinfusion group, the blood samples were collected predose and at 0.25,0.48, 0.58, 0.75, 1.5, 3, 6, 8, 12 and 24 hours after the start ofinfusion. For the oral group, the blood samples were collected predoseand at 0.25, 0.50, 1, 2, 4, 6, 8, 12 and 24 hours after dosing. Theblood samples were collected into Vacutainer™ tubes containing EDTA-K₃as the anti-coagulant and were centrifuged at approximately 4° C. toobtain plasma. The plasma samples were stored at −20° C. until analysisby LC/MS/MS.

A bioanalytical method utilizing high performance liquid chromatographycoupled to tandem mass spectrometry (LC/MS/MS) was developed foranalysis of the selected compound in rat plasma. Detection was performedusing selected reaction monitoring (SRM); Ions representing theprecursor (M+H)⁺ species was selected in quadrupole 1 (Q1) and collidedwith argon gas in the collision cell (Q2) to generate specific production, which was subsequently monitored by quadrupole 3 (Q3). Standardcurve and quality control samples were prepared in male rat plasma andprocessed in the same way as the test samples to generate quantitativedata.

Pharmacokinetic parameters were generated using non-compartmentalpharmacokinetic analysis (Phoenix WinNonlin, version 6.3). Values belowthe lower limit of quantification (LLOQ) were assigned a value of zeroif predose and treated as missing thereafter. Area under the curve (AUC)was calculated using the linear trapezoidal rule. The oralbioavailability (% F) was determined by comparison of the area under thecurve (AUC) of the compound and/or a metabolite generated in plasmafollowing oral administration to that generated following intravenousadministration.

1a 2a 2a J6 2a J6 Example 1b 1a 2a 2a 1a Q30R JFH (t) (s) 2b (t) 2b (s)3a 4a (t) 4a (s) Rat # No. (nM) 1a Q30R JFH J6 2b 3a 4a (nM) (nM) (nM)(nM) (nM) (nM) (nM) (nM) (nM) (nM) % F 422 MC 0.017 C C C B C C C 0.0160.990 0.004 1.155 3.523 0.162 0.375 0.004 0.008 0.028 423 LQ 0.020 C C CC C C C 0.022 0.135 0.004 0.134 0.543 0.012 0.035 0.005 0.008 0.02 426MD 0.033 C C C C C C C 0.035 0.192 0.006 0.073 0.439 0.016 0.038 0.0060.011 0.035 427 ME 0.049 C B C B C C C 0.033 2.514 0.012 3.428 16.0750.198 0.701 0.015 0.016 0.029 434 MG 0.055 C C C C C C C 0.054 0.6630.012 0.337 2.288 0.070 0.010 0.024 0.08 435 MF 0.058 C C C C C C C0.059 0.768 0.012 0.476 2.136 0.083 0.011 0.025 478 478 0.048 C C B C CC 0.045 0.011 3.654 0.820 0.065 0.023 0.019 493 493 0.030 C C A C C C0.036 0.008 5.036 0.650 0.050 0.013 0.018 494 494 0.021 C C C C C C C0.037 0.118 0.005 0.098 0.017 0.005 0.017 0.031 495 495 0.026 C C C C CC C 0.021 0.201 0.008 0.280 1.016 0.135 0.541 0.024 0.014 0.014 498 4980.016 C C C C C C C 0.030 0.040 0.004 0.012 0.039 0.010 0.013 0.0050.011 0.004 499 499 0.032 C C C C C C 0.057 0.008 0.206 0.040 0.0100.021 0.005 503 503 0.041 C C C C C C 0.062 0.012 0.155 0.037 0.0160.028 0.025 507 507 1.503 B C A B C C 3.698 0.297 20.096 1.561 0.3820.594 0.025 510 510 0.050 C C C C C C 0.021 0.009 0.007 0.018 0.0170.020 0.008 0.016 0.072 516 516 0.024 C C C C C C 0.015 0.009 0.0430.013 0.005 0.010 0.025 518 518 0.010 C C B C C C 0.005 0.006 2.4930.643 0.005 0.004 0.011 526 526 0.018 C C C C C C C 0.011 0.095 0.0070.443 1.639 0.132 0.404 0.018 0.011 0.011 536 536 0.013 C C C C C C C0.008 0.048 0.004 0.040 0.154 0.011 0.037 0.009 0.005 0.01 538 LR-10.016 C C C C C C C 0.011 0.015 0.005 0.018 0.048 0.010 0.024 0.0130.010 0.012 539 LT 0.016 C C C C C C C 0.009 0.010 0.004 0.010 0.0470.009 0.039 0.012 0.015 0.009 543 MY 0.015 C C C C C C C 0.016 0.0240.005 0.047 0.165 0.016 0.047 0.010 0.008 0.02 544 MM 0.026 C C C C C CC 0.022 0.044 0.007 0.018 0.064 0.015 0.042 0.020 0.017 0.016 551 OQ0.018 C C C C C C C 0.015 0.102 0.007 0.378 1.492 0.265 0.982 0.0480.020 0.016 552 OR 0.033 C C C C C C C 0.030 0.059 0.011 0.068 0.1600.037 0.109 0.031 0.036 0.006 555 MN 0.021 C C C C C C C 0.014 0.0180.008 0.017 0.071 0.013 0.040 0.034 0.024 0.007 1.5 556 MS 0.008 C C A AC 0.008 0.005 15.564 9.562 0.014 0.02 558 PQ 0.014 C C C C C C 0.0140.106 0.005 0.025 0.155 0.032 0.057 0.015 0.017 561 MP 0.010 C C C C C CC 0.006 0.031 0.004 0.039 0.336 0.032 0.184 0.015 0.015 0.013 24.2 562MO 0.019 C C C C C C C 0.013 0.052 0.007 0.089 0.471 0.137 0.427 0.0510.022 0.018 7.46 563 MT 0.007 C C A A B C 0.003 0.021 44.444 44.4441.092 0.015 0.025 565 NB 0.017 C C C C C C C 0.010 0.034 0.004 0.0200.138 0.007 0.039 0.011 0.023 0.025 566 NC 0.009 C C C C C C C 0.0070.060 0.005 0.089 0.762 0.068 0.191 0.014 0.011 0.026 569 ND 0.024 C C CC C C C 0.020 0.057 0.014 0.314 2.338 0.270 1.319 0.035 0.031 0.044 571OO 0.114 C C C C C C C 0.072 0.139 0.015 0.028 0.259 0.020 0.076 0.0280.086 0.018 572 PF 0.044 C C C C C C C 0.032 0.034 0.014 0.025 0.1270.023 0.117 0.072 0.050 0.018 4.3 573 PG 0.030 C C C C C C C 0.033 0.0430.018 0.045 0.335 0.027 0.119 0.050 0.063 0.008 574 PN 0.016 C C C C C CC 0.015 0.030 0.009 0.027 0.277 0.014 0.035 0.016 0.014 0.02 575 PP0.029 C C C C C C C 0.014 0.043 0.011 0.061 0.403 0.038 0.110 0.0190.027 0.011 576 PO 0.014 C C B C C C 0.013 0.011 3.135 0.470 0.012 0.0130.013 579 OI 0.034 C C C C C C C 0.024 0.086 0.010 0.061 0.326 0.1780.629 0.206 0.036 0.007 580 OJ 0.017 C C C C C C 0.018 0.013 0.502 0.8130.259 0.026 0.017 582 NF 0.010 C C C C C C C 0.011 0.017 0.006 0.0310.218 0.018 0.064 0.017 0.009 0.012 585 NG 0.009 C C C C C C C 0.0060.243 0.010 0.370 1.672 0.063 0.135 0.020 0.008 587 MR 0.041 C C C C C CC 0.028 0.128 0.017 0.079 0.176 0.039 0.053 0.078 0.045 588 588 0.017 CC C C C C C 0.012 0.087 0.004 0.038 0.140 0.012 0.017 0.027 0.019 589 MQ0.011 C C C C C C C 0.008 0.023 0.004 0.017 0.095 0.009 0.023 0.0160.011 590 MU 0.014 C C C C C C C 0.008 0.023 0.004 0.013 0.046 0.0090.025 0.017 0.011 592 NI 0.015 C C C C C C C 0.015 0.059 0.007 0.0260.169 0.019 0.054 0.022 0.020 24.1 594 NP 0.013 C C C C C C C 0.0140.032 0.007 0.010 0.096 0.011 0.043 0.020 0.028 26.1 597 NJ 0.005 C C CC C C C 0.007 0.025 0.007 0.099 0.939 0.094 0.416 0.019 0.017 599 PY0.009 C C C C C C C 0.012 0.013 0.006 0.009 0.098 0.007 0.030 0.0170.018 27.7 600 PZ 0.007 C C C C C C 0.009 0.057 0.005 0.175 1.712 0.0710.386 0.025 601 MV 0.014 C C C C C C C 0.010 0.012 0.004 0.047 0.0340.041 0.011 0.011 602 602 0.025 C C C C C 0.025 0.010 0.053 0.036 0.0360.013 603 603 0.015 C C C C C C C 0.009 0.146 0.007 0.194 0.067 0.0880.015 0.022 604 NR 0.020 C C C C C 0.026 0.006 0.118 0.017 0.017 0.007605 NQ 0.015 C C C C C C 0.017 0.039 0.006 0.064 0.013 0.012 0.013 606OK 0.034 C C C C C C 0.026 0.039 0.011 0.076 0.036 0.045 0.028 607 OL0.067 C C C C 0.047 0.012 0.070 0.052 0.052 608 OH 0.017 C C C C C C0.016 0.031 0.007 0.054 0.023 0.022 0.026 22.7 609 QF 0.005 C C C C C C0.008 0.032 0.004 0.053 0.015 0.011 0.020 610 QA 0.013 C C C C C C 0.0120.045 0.007 0.094 0.052 0.040 0.038 7.46 612 QH 0.005 C C C C C C 0.0070.051 0.005 0.182 0.051 0.049 0.020 613 QG 0.005 C C C C C C 0.005 0.0710.004 0.551 0.065 0.043 0.016 614 OP 0.013 C C C C C C 0.013 0.026 0.0080.018 0.011 0.007 0.027 15.1 615 OM 0.019 C C C C C C 0.014 0.035 0.0090.045 0.053 0.054 0.035 12.2 617 617 0.005 C C C C C C 0.009 0.024 0.0040.032 0.013 0.011 0.019 14.4 618 OT 0.020 C C C C C C 0.011 0.041 0.0050.013 0.012 0.024 619 OF 0.007 C C C C 0.010 0.008 0.445 0.196 620 NK0.081 C C C C 0.049 0.018 0.056 0.154 621 NL 0.013 C C C C 0.011 0.0070.043 0.034 622 NM 0.016 C C C C C C 0.011 0.028 0.006 0.012 0.013 0.0246.93 623 ON 0.006 C C C C 0.005 0.004 0.068 0.086 625 PH 0.031 C C C C CC 0.024 0.051 0.014 0.055 0.042 0.029 626 626 0.030 C C C C C C 0.0210.041 0.009 0.021 0.021 0.027 627 PI 0.039 C C C C C C 0.031 0.094 0.0200.129 0.090 0.065 8.7 628 OG 0.009 C C C C 0.009 0.008 0.601 0.437 631PS 0.006 C C C C A 0.005 0.004 0.077 0.113 44.444 632 PT 0.008 C C C C0.007 0.007 0.383 0.182 633 PR 0.020 C C C C C C 0.013 0.045 0.007 0.0220.028 0.018 634 PU 0.015 C C C C 0.012 0.006 0.068 0.442 635 OU 0.041 CC C C 0.047 0.016 0.040 0.035 636 OV 0.011 C C C C C C 0.010 0.031 0.0070.113 0.046 0.013 637 OW 0.009 C C C C C C 0.009 0.019 0.006 0.009 0.0080.013 13.3 638 OX 0.009 C C C C 0.007 0.006 0.113 0.107 639 QB 0.011 C CC C C C 0.011 0.029 0.008 0.021 0.022 0.028 9.56 640 QE 0.015 C C C C0.013 0.011 0.291 0.563 641 QD 0.030 C C C C 0.025 0.013 0.103 0.193 642QC 0.014 C C C C 0.012 0.008 0.157 0.317 643 643 0.015 C C C C 0.0150.013 0.206 0.607 644 MW 0.026 C C C C C 0.012 0.012 0.020 0.043 0.1230.011 645 MX 0.076 C C C C C 0.036 0.024 0.035 0.070 0.139 0.033 646 6460.109 C C C C C 0.058 0.030 0.042 0.112 0.262 0.034 648 PJ 0.088 C C C CC 0.068 0.058 0.136 0.335 0.854 0.056

1. A method of treating hepatitis C virus, said method comprisingadministering to a human patient in need thereof, a pharmaceuticalcomposition which comprises a therapeutically effective amount of: (1) acompound of formula:

(2) a NSSB polymerase inhibitor; and (3) a NS3 protease inhibitor. 2.The method of claim 1, wherein the NS3 protease inhibitor is boceprevir.3. The method of claim 1, wherein the NS3 protease inhibitor istelaprevir.
 4. The method of claim 1, wherein the NS3 protease inhibitoris TMC435350.
 5. The method of claim 1, wherein the NS3 proteaseinhibitor is BI-1335.
 6. The method of claim 1, wherein the NS3 proteaseinhibitor is BI-1230.
 7. The method of claim 1, wherein the NS3 proteaseinhibitor is MK-7009.
 8. The method of claim 1, wherein the NS3 proteaseinhibitor is VBY-376.
 9. The method of claim 1, wherein the NS3 proteaseinhibitor is ITMN-191.
 10. The method of claim 1, wherein the compoundof formula:

and the NSSB polymerase inhibitor are administered together with the NS3inhibitor.
 11. The method of claim 1, wherein the compound of formula:

and the NS5B polymerase inhibitor are administered separately from theNS3 inhibitor.